| A 25 year-old female presented in the emergency room with an acute urethral discharge of 2 days duration. A smear for gram stain was obtained (see accompanying image). Many polymorphonuclear leukocytes and intracellular and extracellular gram negative diplococci were observed. Based on the clinical history and the gram stain observation, a diagnosis of gonorrhea can be made. | View Page |
| Review 2 Smith KR, Fisher HC III, Hook, EW III: Prevalence of fluorescent monoclonal antibody-nonreactive Neisseria gonorrhoeae in five North American sexually transmitted disease clinics.J Clin Microbiol 34:1551-1552, 1996We compared a direct fluorescent monoclonal antibody (DFA) test with alternative enzymatic and fermention tests for identifying presumptive gonococcal isolates in a systematic sample from patients attending five sexually transmitted disease clinics in five cities.Fourteen (2.5%) of 556 isolates from three clinics were nonreactive with the DFA confirmatory reagent and reactive by both the Quad-Ferm and Rapid NH tests. The prevalence of DFA-nonreactive Neisseria gonorrhoeae isolates varies geographically and is independent of local methods for the identification of possible gonococci.On the basis of our findings, we recommend that for use in medicolegal and other instances in which a diagnosis of gonorrhea has the potential to have far-reaching effects, it is appropriate to test DFA reagent-nonreactive, oxidase-positive, gram-negative diplococci by alternative methods of gonococcal confirmation.Although the prevalence of such isolates could change, the fluorescent monoclonal antibody confirmation reagents remain useful for many clinical situations. Their ease of use and ready applicability for screening large numbers of isolates make them useful for many laboratories. | View Page |
| The recovery of an oxidase-positive, gram-negative diplococcus that tests DFA-nonreactive should be tested by alternative methods when establishing a fool-proof diagnosis of gonorrhea. | View Page |
| The most important modifiable risk factor for enteric colonization with vancomycin-resistant Enterococcus faecium is: | View Page |
| Clinical History The prototype history for this organism is either a still birth or a neonate with death ensuing within 2 or 3 days post-partem due to high fever, sepsis, and respiratory distress. The mother usually experienced a flu-like illness late in the third trimester of pregnancy, characterized by low-grade fever, myalgias, malaise and backache. In this case, biopsy material of brain tissue obtained at autopsy was submitted to the pathology laboratory for tissue diagnosis and fluid from the pia-arachnoid was sent to the microbiology laboratory for culture. | View Page |
| Histology of Brain Biopsy The H & E section of the brain biopsy (left frame)revealed edema of the parencymya with the accumulation of inflammatory cells in the perivascular spaces. The close in view of the exudate (right) frame reveals that the inflammatory exudate is comprised primarily of polymorphonuclear luekocytes. The histologic diagnosis therefore is suppurative meningitis, with culture results necessary to establish the etiologic agent. | View Page |
| Introduction Hereditary hemochromatosis (HH) is a disorder of iron regulation that results in excessive dietary iron absorption through the gastrointestinal tract. Over time, the resultant iron overload and its deposition in tissue may lead to widespread organ damage, a variety of chronic disorders, and even death. Although it is a genetic disorder, clinical symptoms most typically become apparent in middle aged adults. Iron overload occurs in a variety of hereditary and acquired forms, known as iron storage diseases. HH is the most common cause of inherited iron overload. (1) Due to lack of awareness, HH often goes undetected or unrecognized by health care providers. Early detection to prevent the serious complications associated with iron overload has important consequences for reducing morbidity and mortality. Laboratory tests that assess iron levels and molecular assays for genetic mutatations are essential for both its detection and diagnosis. | View Page |
| References 1. Beutler E. Iron storage disease: Facts, fiction and progress. Blood Cells Mol Dis. 2007;39:140-7.2. Higgins T, Beutler E, Doumas BT. Hemoglobin, iron, and bilirubin. In: Burtis CA, editor. Teitz Fundamentals of Clinical Chemistry. 6th ed. Saunders Elsevier, 2008.3. Ganz T. Hepcidin, a key regulator of iron metabolism and mediator of anemia and inflammation. Blood 2003;102(3):78-8.4. Andrews NC, Schmidt PJ. Iron homeostasis. Annu Rev Physiolo. 2007;69:69-85.5. Murtagh LJ, Whiley M, Wilson S, et al. Unsaturated iron binding capacity and transferrin saturation are equally reliable in detection of HFE hemochromatosis. Am J Gastroenterol. 2002;97(8):2093-9.6. Haddy TB, Castro OL, Rana SR. Hereditary hemochromatosis in children, adolescents, and young adults. Am J Pediatr Hematol Oncol 1988;10:23-4.7. Edwards CQ, Ajoika RS, Kushner JP. Hemochromatosis: A genetic definition. In Barton JC, Edwards CQ, eds. Hemochromatosis: Genetics, Pathophysiology, Diagnosis and Treatment. Cambridge, UK:Cambridge Univ Pr 2000:8-11.8. Whitlock EP, Garlitz BA, Harris EL , et al. Screening for Hereditary Hemochromatosis: A Systematic Review for the U.S. Preventive Services Task Force. Ann Intern Med. 2006; 145: 209-23.9. Wallace DF, Subramaniam VN. Non-HFE haemaochromatosis. World J Gastroenterol. 2007;13(35):4690-8.10. Tavill AS. Diagnosis and management of hemochromatosis. Hepatology. 2001;33:1321-811. Qaseem A, Aronson M, Fitterman N, Snow V, Weiss KB, Owens DK, et al. Screening for hereditary hemochromatosis: a clinical practice guideline from the American College of Physicians. Ann Intern Med. 2005;143:517-21.12. Phatak PD, Bonkovsky HL, and Kowdley KV. Hereditary Hemochromatosis: time for targeted screening. Ann Intern Med. 2008; 149(4): 270 – 2.13. Brissot P, deBels F. Current approaches to the management of hemochromatosis. Hematology Am Soc Hematol Educ Program. 2006:36-41. 14. Guidance for industry: Variances for blood collection from individuals with hereditary hemochromatosis. http://www.fda.gov/cber/gdlns/hemchrom.htm Accessed 12/17/08. | View Page |
| General Clinical Considerations Hereditary hemochromatosis (HH) is frequently discovered only during management of associated illness or routine health evaluations. It has been estimated that only a small percentage of all affected persons are actually diagnosed. Individuals with HH may be symptomatic for several years prior to diagnosis and may have consulted multiple health care providers.Under-diagnosis of HH is thought to occur due to:• Lack of specificity of early signs and symptoms• Asymptomatic status of some patients until damage to organs and tissues has occurred• Confusion with liver disease due to other causes• Insufficient awareness and knowledge of HHEarly identification of persons with HH is essential to prevent serious and irreversible complications associated with severe iron overload. A classic triad of skin hyperpigmentation (bronzing), type 2 diabetes, and hepatic cirrhosis has long been recognized as evidence of advanced iron overload. However, persons with HH may present with a much wider variety of signs and symptoms, particularly if they are seen before significant iron accumulation has occurred. Age of presentation and disease severity are highly variable. A diagnosis of HH is based on laboratory evidence of iron overload, genetic mutations associated with HH, and presence of clinical signs and symptoms consistent with HH.(10) | View Page |
| Secondary Disorders of Iron Overload In addition to hereditary hemochromatosis (HH), there are other conditions of iron overload that must be considered in a differential diagnosis. Disorders such as sickle cell disease, thalassemia, sideroblastic anemia, congenital dyserythropoietic anemia, and liver disease may also cause iron overload. Transfusion-dependant patients and persons who abuse iron-containing vitamin supplements are also at risk. These conditions are usually described as secondary iron overload, in contrast to the primary iron overload of HH.Patient history, clinical signs and symptoms, biochemical and hematologic laboratory analyses, and possibly results of a liver biopsy may be needed to establish a diagnosis of a condition causing secondary iron overload. DNA tests for common HFE mutations are very likely the most important diagnostic tool for identifying HH as the cause of iron overload. In some patients, both secondary causes and HH may be contributing to iron overload. Differentiating the secondary causes of iron overload from HH is heavily dependent on the results of laboratory assays, but a complete discussion is beyond the scope of this course. | View Page |
| Diagnosing HH The diagnosis of hereditary hemochromatosis (HH) is made through a combination of laboratory tests and medical evaluation of a patient's signs and symptoms. Iron overload is identified by tests that evaluate iron metabolism, while molecular assays are needed to document mutations in the HFE gene or others such as hepcidin, hemojuvelin, or transferrin receptor. Individuals with documented iron overload who exhibit signs and symptoms consistent with HH and who possess HFE or other mutations are considered to have HH. Other causes of secondary iron overload may need to be ruled out.An example of a testing algorithm is shown. | View Page |
| Which of the following is (are) needed for a diagnosis of hereditary hemochromatosis (HH)? | View Page |
| Prognosis and Mortality The major determinant of prognosis in cases of hereditary hemochromatosis (HH) is the degree of organ damage from iron overload at the point of diagnosis. The presence of liver cirrhosis reduces life expectancy. Damage that has occurred to tissues and organs is irreversible, but further damage can be halted with treatment. When there is no evidence of cirrhosis at time of diagnosis, life expectancy may be equal to that of persons without HH. With proper management of HH through treatment, affected individuals have good long-term outcomes. Hepatocellular carcinoma associated with cirrhosis, hepatic failure, and cardiac failure are the most common causes of death in persons with HH. Compared to the normal population, liver cancer is many times more prevalent as a cause of death in persons with HH. Cardiomyopathy, diabetes, and cirrhosis are all more common causes of death among persons with HH than among normal persons. The earlier HH is detected, before the onset of severe organ damage, the lower the risk of mortality. | View Page |
| Transferrin and Total Iron Binding Capacity The test for transferrin (Tf) measures the concentration of the primary carrier protein for iron. Measuring total iron binding capacity (TIBC) is an indirect method of assessing transferrin and provides comparable information. The TIBC (or transferrin) are typically performed along with the SI. Taken together, these determinations are useful in the differential diagnosis of many disorders affecting iron metabolism, including hereditary hemochromatosis (HH) and iron deficiency anemia. Tf and TIBC are typically low-normal or decreased in HH and are increased in iron deficiency. Serum transferrin can be measured directly using immunochemical methods such as nephelometry and turbidimetry. TIBC is performed in a 2-step method by adding ferric iron to the specimen in sufficient quantity to completely fill all of the iron binding sites on transferrin. Excess, unbound iron is removed by adsorption with magnesium carbonate, alumina, or ion resin. The iron content of the saturated binding protein is then measured as described for SI. Serum is the specimen of choice for Tf and TIBC. TIBC is less subject than SI to day-to-day variation and other causes of variability.A typical reference interval for TIBC is 300 - 360 micrograms/dL.(2) | View Page |
| Serum Ferritin Serum ferritin (SF) level reflects the amount of storage iron in tissues. An elevated SF combined with elevated TS implies primary iron overload. Patients with hereditary hemochromatosis (HH) generally show increases in SF as adults, but a normal SF does not rule out the diagnosis of the disease. Children and premenopausal females with HFE mutations may have had inadequate time to develop iron overload, but may do so later in life.SF alone is inadequate as the sole screening test for HH because it lacks the necessary sensitivity and specificity. SF is frequently elevated in persons with inflammation, cancer, or infection. SF is often ordered along with the serum iron and TIBC when iron overload is suspected. SF is also important is assessing the efficacy of treatment of HH.Upper limits of reference intervals for SF are 200 ng/mL for premenopausal women and 300 ng/mL for men and postmenopausal women. 40 ng/mL is a typical lower limit for the reference interval.SF is measured in serum using immunochemical methods such as enzyme-linked immunosorbent assay (ELISA), immunoradiometric assay, immunochemiluminescent assay, and immunofluorometry. SF tests are available as automated assays and in kit form.(2) | View Page |
| Screening Controversies The subject of screening for hereditary hemochromatosis (HH) is controversial and is currently being debated in the medical literature. Using laboratory tests to screen the asymptomatic general population is currently not recommended due to issues of testing costs, low genetic penetrance, and the possible risk of discrimination. Targeted case finding in select high risk populations such as men of Northern European ancestry may be a better approach to screening. (12)Molecular-based (DNA) assays required for confirmation of HH are costly when used for general population screening. Because recent studies have shown that a high percentage of persons with C282Y mutations do not develop iron overload or HH-related clinical conditions, screening for these mutations may falsely label an individual with a disease diagnosis. At the present time, it is impossible to determine which homozygotes or heterozygotes for HFE mutations will eventually develop iron overload. Furthermore, there is potential risk of discrimination in obtaining health insurance for persons identified as having genetic disorders.In contrast, some experts do advocate for screening the general population. Mutations associated with HH are very common in Caucasians in the US. Individuals who know they carry mutations associated with HH may benefit from periodic testing for iron overload. Finally, laboratory tests that assess iron status are relatively inexpensive, widely available, and offer one approach to screening for phenotypic expression of HH. Screening first-degree family members of a person with documented HH is generally considered to be worthwhile. Early detection of HH in relatives with common mutations may permit treatment before the development of substantial iron overload and related disease due to organ damage. | View Page |
| What is one established reason supporting general population screening for hereditary hemochromatosis (HH)? | View Page |
| Molecular Tests DNA tests for HFE mutations associated with hereditary hemochromatosis (HH) are available in some clinical laboratories and reference laboratories. Testing for the presence of the C282Y is essential, although most labs also test for H63D and S65C mutations. Molecular testing is most appropriate for confirmatory testing of symptomatic individuals with altered iron studies (increased TS and SF), in pre-symptomatic individuals (increased TS, normal SF and liver function tests), and in family members of individuals diagnosed with HH. The use of genetic tests alone for routine screening of asymptomatic persons is not recommended for several reasons. A positive test indicating the presence of HFE mutations does not guarantee that an individual will develop clinically significant iron overload or predict severity of symptoms. A negative result (no HFE mutations present) does not rule out a diagnosis of iron overload because of genetic heterogeneity. Compared to biochemical analyses for iron, molecular assays are expensive. Finally, molecular testing may result in the diagnosis of a genetic disease, thus opening up the possibility for discrimination in health insurance coverage. Using molecular methods, DNA is extracted from leukocytes in whole blood samples or from buccal cells and analyzed for specific HFE mutations using polymerase chain reaction (PCR) with melt curve analysis. Currently there are no FDA-cleared products for HFE testing, and testing laboratories are using "home brew" reagents. This situation is expected to change as manufacturers submit products for FDA approval. | View Page |
| Which statement best describes the use of the liver biopsy in suspected cases of hereditary hemochromatosis (HH)? | View Page |
| Billing and medical necessity Billing:
Highest risk activity a laboratory has. All laboratory activities contribute to the billing process.
Many of the risk areas included in this program are components of the billing function.
Medical necessity:
Medicare is only allowed, by law, to pay for tests that are reasonable and necessary for the diagnosis and treatment of disease.
Medical necessity is an underlying principle of the Medicare program.
Tests performed for screening or routine exams are not considered medically necessary by the Medicare program.
| View Page |
| Medical coverage policies (LMRPs) LMRPs (Local Medical Review Policies) are published by Medicare for some laboratory tests.
Developed for tests that can be used for screening or diagnosis of disease.
CPT codes describe laboratory tests and ICD-9CM codes determine when coverage is allowed.
If an LMRP test is ordered by a physician, an ICD-9CM code that is included in the LMRP must be given to the laboratory or the Medicare program will not pay for the test.
It is against the law for laboratory to change or add an ICD-9 code submitted by a physician.
The Balanced Budget Act of 1997 made it illegal for physicians to order LMRP tests and not supply an ICD-9CM code with the order.
| View Page |
| Requisitions and ambiguous orders Requisitions must be designed to ensure that ordering physicians can choose tests that are medically necessary for their patients.
Requisitions should contain reminders about Medicare rules of medical necessity and list the contents of panels and profiles.
Requisitions must provide a place for the physician to include diagnosis (ICD9-CM) codes.
Physicians should be encouraged to use only the requisitions supplied by the laboratory to order tests.
Ambiguous or unclear test orders
When the orders for a test are not absolutely clear, the laboratory must contact the ordering physician to clarify the orders before performing and billing for the test.
The laboratory cannot guess at the order.
The laboratory cannot perform and bill for tests that are not specifically ordered.
The laboratory cannot change a physician order without contacting the physician.
In any case where specimen integrity or patient care will be compromised by a delay in testing follow the policies the laboratory has established for such cases.
| View Page |
| Medical necessity Medical necessity means that Medicare is not allowed by law to pay for any tests that are not necessary for diagnosis or treatment of disease.A laboratory may not submit a claim to Medicare or other government payers for any test it knows is not medically necessary except in certain cases:
When the patient has signed an advance notice.
When a patient has requested the lab to submit such a claim for a determination by Medicare.
Medicare does not pay for screening tests or tests that are ordered in the absence of signs or symptoms.Billing department employees are responsible to follow all policies and procedures related to the submission of claims to reduce erroneous billings. | View Page |
| Local medical review policies (LMRPs) LMRPs (Local Medical Review Policies) are published by Medicare for some laboratory tests.
They are usually developed for tests that can be used for screening or diagnosis of disease.
LMRPs use CPT codes to identify the tests and ICD-9 codes to determine when coverage is allowed.
If an LMRP test is ordered by a physician, an ICD-9 code that is included in the LMRP must be given to the laboratory or the Medicare program will not pay for the test.
It is against the law for laboratory to change or add an ICD-9 code submitted by a physician.
A laboratory should not submit a claim for an LMRP test that is not accompanied by an acceptable ICD-9 code.
The Balanced Budget Act of 1997 made it illegal for physicians to order LMRP tests and not supply an ICD-9CM code with the order.
| View Page |
| Documentation All information related to diagnosis or other billing information received from a physician office must be documented.
Documentation includes the name of the person collecting the information, the name of the person giving the information, and the date.
This documentation must be linked to the original order.
Billing department employees must ensure that complete records and documentation exist for all billing transactions.Not documented means not done.All communication, (either written or verbal), with government, Carrier, or Fiscal Intermediary representatives must be documented.Employees should report instances where records are missing, incomplete, or improperly filed, to ensure that corrective action is taken. | View Page |
| Communication with physicians and patients It is important that billing department employees are clear and accurate when communicating with physician office personnel and patients.
Never guess at the answer to a question; ask if you are unsure.
Do not speculate or express personal opinions.
When requesting diagnosis information from the physician office staff be careful to not lead them to give a billable code:
The code must come from the patient's medical record.
There is an incentive program for patients to find and report fraud and abuse by health care providers, including laboratories, so:
Billing department employees must accurately state laboratory policies and procedures, or forward the call to a supervisor to avoid misstatements and misunderstandings.
| View Page |
| Case Study 1 A billing clerk is entering billing demographics on requisitions as a part of the normal days work. The department is under pressure to reduce accounts receivable, which means that the more clean claims that are filed, the better. This particular requisition is for a Medicare patient and has an LMRP test but does not have a diagnosis on it. She remembers that just a few requisitions before this one she had a requisition from the same doctor that had this same test on it that did have a diagnosis that allowed the test to be billed. She thumbs back in the pile and finds the previous requisition, notes the code that was used and adds it to the current requisition. This will help her meet the department goal of getting claims paid and reducing accounts receivable. It is all right for her to do this because:Correct Answer: She should not do this because it is against the law to change diagnosis information on a requisition.Discussion:
A laboratory employee should never change, add or use previously received diagnosis information for the purpose of making a test billable for the Medicare program or for any other insurance or payer. This is a form of fraud and for each claim submitted as a result of this activity the laboratory is liable for a false claim and would have to pay the government back three up to times the reimbursement for the test and up to $10,000 for each claim submitted. Further, if the employee is caught doing this, even if the employee is ignorant of the law and any laboratory policy prohibiting it, she must be disciplined and so should the supervisor. Any employee who notices another employee doing this should correct the employee and report the incident to the department supervisor immediately. | View Page |
| Test Requisitions Requisitions must be designed to ensure that ordering physicians can choose tests that are medically necessary for their patients. Requisitions should contain reminders about Medicare rules of medical necessity and list the contents of panels and profiles. Requisitions must provide a place for the physician to include diagnosis (ICD9-CM) codes. Physicians should be encouraged to use only the requisitions supplied by the laboratory to order tests. | View Page |
| Case Study 1 A billing clerk is entering billing demographics on requisitions as a part of the normal days work. The department is under pressure to reduce accounts receivable, which means that the more clean claims that are filed, the better. This particular requisition is for a Medicare patient and has an LMRP test but does not have a diagnosis on it. She remembers that just a few requisitions before this one she had a requisition from the same doctor that had this same test on it that did have a diagnosis that allowed the test to be billed. She thumbs back in the pile and finds the previous requisition, notes the code that was used and adds it to the current requisition. This will help her meet the department goal of getting claims paid and reducing accounts receivable. Can she do this?Correct Answer: She should not do this because it is against the law to change diagnosis information on a requisition.Discussion: A laboratory employee should never change, add or use previously received diagnosis information for the purpose of making a test billable for the Medicare program or for any other insurance or payer. This is a form of fraud and for each claim submitted as a result of this activity, the laboratory is liable for a false claim and would have to pay the government back up to three times the reimbursement for the test and up to $10,000 for each claim submitted. If the employee is caught doing this, even if the employee is ignorant of the law and any laboratory policy prohibiting it, she must be disciplined along with her supervisor. Any employee who notices another employee doing this should correct the employee and report the incident to the department supervisor immediately. | View Page |
| Local medical review policies (LMRPs) Local Medical Review Policies (LMRPs) are published by Medicare for some laboratory tests. They are usually developed for tests that can be used for screening or diagnosis of disease. LMRPs use CPT codes to identify the tests and ICD-9 codes to determine when coverage is allowed. If an LMRP test is ordered by a physician, an ICD-9 code that is included in the LMRP must be given to the laboratory or the Medicare program will not pay for the test. It is against the law for a laboratory to change or add an ICD-9 code submitted by a physician. A laboratory should not submit a claim for an LMRP test that is not accompanied by an acceptable ICD-9 code. The Balanced Budget Act of 1997 made it illegal for physicians to order LMRP tests and not supply an ICD-9CM code with the order. | View Page |
| Medical necessity The Centers for Medicare and Medicaid Services (CMS), the US agency that administers the Medicare program, defines "medical necessity" as services or items reasonable and necessary for the diagnosis or treatment of illness or injury or to improve the functioning of a malformed body member. Medicare will not pay for any tests that CMS determines as unnecessary for diagnosis or treatment of disease.A laboratory may not submit a claim to Medicare or other government payers for any test it suspects is not medically necessary unless: The patient has signed an Advanced Beneficiary Notice, or A patient has requested the lab to submit such a claim for a determination by Medicare. Medicare does not pay for screening tests or tests that are ordered in the absence of signs or symptoms. Billing department employees are responsible for following all policies and procedures related to the submission of claims to reduce erroneous billings. | View Page |
| Laboratory Billing Department Communication With Physicians and Patients It is important that billing department employees are clear and accurate when communicating with physician office personnel and patients. Never guess at the answer to a question; ask if you are unsure. Do not speculate or express personal opinions. When requesting diagnosis information from the physician office staff, be careful to not lead them to give a billable code. The code must come from the patient's medical record. Billing department employees must accurately state laboratory policies and procedures, or forward the call to a supervisor to avoid misstatements and misunderstandings. | View Page |
| Documentation All information related to diagnosis or other billing information received from a physician office must be documented. Documentation includes the name of the person collecting the information, the name of the person giving the information, and the date. This documentation must be linked to the original order. Billing department employees must ensure that complete records and documentation exist for all billing transactions. Not documented means not done. All communication, (either written or verbal), with government, Carrier, or Fiscal Intermediary representatives must be documented. Employees should report instances where records are missing, incomplete, or improperly filed, to ensure that corrective action is taken. | View Page |
| An 8 year old girl is protected from severe hemolytic anemia by an elevated fetal hemoglobin level ( hemoglobin F). | View Page |
| DIC: graft vs. host disease The peripheral smear illustrated in the photograph was obtained from a patient with a recent renal transplant. The patient developed a rash, accompanied by nausea and diarrhea. Graft vs. host disease was clinically suspected. The peripheral smear findings are consistent with that diagnosis. The presence of spherocytes suggests a hemolytic process which is supported by the presence of nucleated RBCs. A few scattered schistocytes and the decrease of platelets suggests DIC. The presence of target cells presents the possibility of associated liver disease. Additional tests, particularly coagulation studies, should confirm the diagnosis of microangiopathic hemolytic anemia. | View Page |
| Hemolytic disease of the newborn Jaundice was recognized in a day-old infant. Notice particularly the size variation (anisocytosis) of the erythrocytes on the infant's peripheral smear. What does this observation mean? Does it provide immediate information that might serve as guidance in expediting diagnosis and treatment? Note that normal-sized red blood cells, microcytes, microspherocytes, macrocytes, and nucleated red blood cells are all present. Red cell variations are expected findings in healthy neonates, but the variations here are exaggerated. Hyposplenic functional features may appear, including acanthocytes, spherocytes, and possibly Howell-Jolly bodies, especially if hemolysis is particularly vigorous. A high (3-7%) reticulocyte count is not unusual during the first three or four days after birth, however, the marrow in this jaundiced infant is proliferating vigorously in response to hemolysis. A call for more red cells is urgent. Immature red cells (in the form of nucleated red cells) and red cells with stippling of RNA (basophilic stippling) are readily identified. Red cell maturation sequence has not been totally processed in the marrow nor is all residual red cell debris removed by the spleen. In the lower photograph are reticulocytes stained by supravital stain (new methylene blue). Basophilic stippling (specks of RNA) stains with both supravital stains and with routine Wright-Giemsa stain. | View Page |
| Considering the predominance of microspherocytes on the blood smear, and the patient's jaundiced condition, what is the most likely diagnosis? | View Page |
| Atypical smear: Case follow-up The patient whose blood smear is shown in the photograph was a 32-year-old female from Virginia who came to the high country of Colorado to ski. The day after arrival, she experienced shortness of breath, fatigue, and upper abdominal pain. She was seen in a medical center in the mountains where a working diagnosis of altitude sickness was made. A CBC revealed RBCs 5.1 x 1012/L, hemoglobin 12.8g/dL, MCV 60fL, hematocrit 40.9%, and normal total WBC, differential, and platelet count. The RDW was normal. Further questioning revealed a previous diagnosis of heterozygous beta-chain thalassemia. No other abnormal hemoglobins were found on hemoglobin electrophoresis, but HbA-2 was elevated to 5%, supporting the diagnosis of beta thalassemia. The patient's poikylocytosis and anisocytosis may be a clue to an underlying erythrocyte abnormality. Persons with iron deficiency anemia may experience various degrees of hypoxia upon arriving at high altitudes. Those with sickle cell disease and thalassemia minor (as in this case) may experience bone pain or other symptoms of "crisis" and/or alteration in the appearance of their erythrocytes upon sudden high altitude exposure. The classic teaching is that in differentiating iron deficiency anemia from thalassemia, increased RDW would favor iron deficiency; normal RDW favors thalassemia. | View Page |
| The photograph is representative of the peripheral blood smear of a five-month-old immigrant from Asia. Her mother was concerned that the child was not eating well. Her spleen was palpable.The hemogram revealed the following:Hb 9.6g/dL (normal 12.0 - 16.0 g/dL)RBC 5.48 X 1012/L (normal 4.2 - 5.9 X 1012/LHCT 30.4% (normal 37 - 48%)MCV 55.4 fl (normal 86 - 98 fl)MCH 17.5 pg (normal 27 - 32 pg)MCHC 31.6 g/dL (normal 31 - 37 g/dL)RDW 34.9% (normal 11 - 15%)Reticulocyte count 10.9% (normal 0.5 - 1.5%)Select the most likely diagnosis based on the clinical information and peripheral blood findings. | View Page |
| Hb E disease (continued) The family (cited in the previous case history) was from a region of Thailand where the physician knew HbE carriers are prevalent. Homozygous hemoglobin E is common in Southeast Asia and presents with very mild anemia and seldom requires transfusion. Over 30 million people in the world are HbE carriers, making this abnormal hemoglobin almost as common as HbS. Hemoglobin E is uncommon in North America and in Europe, but with changing immigration patterns, hemoglobinopathy E cannot be ignored. Peripheral blood smear findings of target cells, microspherocytes, red cell hypochromia, a few red blood cell fragments, and nucleated red blood cells require evidence from hemoglobin electrophoresis to establish a diagnosis. Clinically, a very important and severe syndrome is hemoglobin E/beta thalassemia in which there is hemolysis requiring repeated transfusions. The patient has a severe anemia, low MCV (50's), and high RBC. This is characteristic of Hgb E/beta thalassemia. | View Page |
| The patient, an 8-month-old girl, was anemic, jaundiced, and had splenomegaly. Her family had immigrated from the Middle East. Based on the history and the peripheral blood picture, the most probable diagnosis is thalassemia. | View Page |
| Reporting of laboratory data in regard to blood cell abnormalities Laboratory data must be presented to clinicians in a user friendly way to promote effective decision making. Databases must be designed to provide clear information that leads quickly to the best patient care outcome. We continue learning how to collect and retrieve laboratory data from our machines, but we are not always in tune to how entry and retrieval of data is geared to and, more directly, influences patient care outcomes. Examples of blood cell abnormalities on a peripheral blood smear that may immediately direct the physician to a specific diagnosis are: (1) presence of target cells as found in thalassemia or hemoglobinopathies and target cells in liver disease, particularly with obstructive jaundice; (2) burr cells as a signal of chronic renal disease and uremia; and (3)atypical neutrophil inclusions relating to genetic disorders. Critical appraisal of such observations could add valuable clues for a diagnosis. Laboratory professionals must establish a set of principles for orderly observation of blood cell morphology, have a clear vision of the applications of their work, and understand the potential clinical implications of their reports and interpretations. Emphasis on values and relevance focuses on patient care outcomes and their dependency on prompt availability of results and contextual interpretations. | View Page |
| Case History A 54 year-old man was brought to the clinic by his sister who was emphatic that her brother was "not taking care of himself."The patient had a previous gastrectomy and splenectomy. He also had a diagnosis of alcoholism, malnutrition, and hepatic cirrhosis. The following five pages discuss a variety of erythrocyte changes that have occurred as a result of his various conditions. | View Page |
| Conditions Associated with Spherocytes Examples of conditions in which spherocytes can be seen include hereditary spherocytosis and immune hemolytic anemias (i.e., ABO incompatibility). Spherocytes can also form in conditions where there has been a direct physical or chemical injury to the cells. An example would be a smear from an individual who has suffered severe burns. In hereditary spherocytosis, a condition where spherocytes are numerous, the MCHC value will be at the upper limits of normal, or about 36. The identification of spherocytes on the smear of a patient with hereditary spherocytosis can aid significantly in the diagnosis of the disorder. In Artifactual spherocytes can appear when blood is stored for a prolonged period of time. | View Page |
| Examples of Stomatocytes Examples of stomatocytes can be seen in the center of this slide and to the left of the center. Conditions in which a significant number of in vivo stomatocytes can be seen include hereditary stomatocytosis, neoplastic disorders, liver disease and Rh null disease. The largest numbers of in vivo stomatocytes are seen in hereditary stomatocytosis and their identification is necessary to make the diagnosis. | View Page |
| Summary It is important to differentiate in vitro changes which are secondary to preparing the slide, from in vivo morphology, which is the result of the pathophysiological condition of the patient. Examining erythrocytes in the critical viewing area is extremely important in making this distinction. The determination of the clinical significance of the morphology reported is the responsibility of the physician, who must correlate the blood smear findings with the clinical diagnosis, and other laboratory parameters. | View Page |
| The identification of which of the following abnormal forms may contribute significantly to specific clinical diagnosis: | View Page |
| Another Echinocyte Another example of an echinocyte is seen in the center of this slide. In rare instances, echinocytes circulate in vivo in uremia, following heparin injection, in certain congenital anemias and in pyruvate kinase deficiency. Plastic slides must be used to verify the presence of in vivo echinocytes. Since echinocytes do not aid in the diagnosis of these conditions, their main importance lies in the fact that they are artifactual and reversible and must be distinguished from acanthocytes. | View Page |
| Sickle Cell Anemia Sickle cells can be seen in the peripheral blood of patients who have homozygous sickle cell anemia; however other tests are needed to make the diagnosis. Most sickled cells can revert back to the discoid shape when oxygenated. About 10% of sickled cells are unable to revert back to their original shape after repeated sickling episodes. | View Page |
| Another Target Cell Another example of a target cell (or codocyte) is seen in the center of this slide. Notice that the hemoglobin in the center of this cell is somewhat lighter in appearance than in the previous slide. A second codocyte can be seen in the upper left portion of the slide. Codocytes appear in conditions which cause the surface of the red cell to increase disproportionately to its volume. This may result from a decrease in hemoglobin, as in iron deficiency anemia, or an increase in cell membrane.
Target cells have excess membrane cholesterol and phospholipid and decreased cellular hemoglobin. Examples of other conditions in which target cells may be present include thalassemias, hgb C disease, post splenectomy and obstructive jaundice. Since their presence can be the result of an in vitro artifact, their value in clinical diagnosis is limited. | View Page |
| Another Keratocyte Another example of a keratocyte (helmet cell) is seen in the center of this field. Examples of conditions in which keratocytes can be seen include intravascular coagulation, microangiopathic hemolytic anemia, glomerulonephritis, and rejection of renal transplants. The diagnosis of these disorders is not based on the presence of keratocytes. | View Page |
| Identification of white cell morphological changes are important because: | View Page |
| Importance of Recognition It is important to be able to recognize the presence of these changes and then identify them for several reasons:If the changes are pathological, their identification may aid the physician in diagnosing a specific condition.If the changes are not pathological, their identification alerts the physician to the fact that the changes are present, thus avoiding a possible misdiagnosis.If reactive, it indicates that although the cells are functioning normally, they are reacting to a stimulus. Indicating the presence of such cells may aid in determining the diagnosis or monitoring the course of disease once a diagnosis has been made. | View Page |
| All of the following are important reasons to identify cell changes EXCEPT: | View Page |
| Normal Band Forms vs. Pelger-Huet Bands Recognition and diagnosis of the inherited form is important because many of these Pelger-Huet neutrophils may be classified as bands, therefore; increased numbers of bands might be erroneously reported in these patients. Since increased bands frequently indicate infection, reporting Pelger-Huet cells as normal band forms could result in inappropriate treatment for infection. Pelger-Huet cells have denser nuclear chromatin than neutrophilic band forms. | View Page |
| More on Dohle Bodies Dohle bodies are seen in a number of conditions, including infections, burns, measles, leukemia and chemotherapy. Dohle bodies are classified as pathological in the sense that they are only present when the body is responding to an unusually severe stress or stimulus. This severe stress may cause the cytoplasm of some cells to mature improperly. Their presence does not aid in the diagnosis of the disorders in which they are found, but they are frequently seen along with toxic granulation and/or vacuoles often present in infections and burns. Recognition is important because their appearance is similar to May-Hegglin bodies, which appear in a rare hereditary disorder called May-Hegglin anomaly. | View Page |
| Alder-Reilly Neutrophil An example of a normal neutrophil, lower left, and one showing some increased granulation typical of that seen in Alder-Reilly anomaly. Morphologically, it may be difficult to distinguish these granules from toxic granulation, however, the diagnosis is made on the basis of the presence of the many distinctive physical characteristics. | View Page |
| Additional comments on this exercise The following pages in this presentation includes a series of white blood cell abnormalities that may be identified in a peripheral blood smear. Many of the cases will simulate the practice of a peripheral smear review by a hematology morphologist. He/she must asses what responses in patient care may be triggered by the clinician attempting to interpret the reported findings on a peripheral smearObservations of white blood cell abnormalities in the peripheral blood smear should be reported so as to direct the physician to an immediate specific diagnosis, such as: (1) atypical lymphocytes suggesting infectious mononucleosis rather than leukemia, (2) toxic granules in neutrophils as in acute infections, or atypical granules suggesting a genetic disorder, (3) an unusual mix of cells, such as too many or too few neutrophils, monocytes, or other myeloid cells, and (4) the presence of giant platelets, myelocytes, or other cells suggesting a myelodysplastic syndrome.In summary, laboratory data should be presented to clinicians in a user friendly way to promote effective decision making. The design of the data base of information must be directed toward providing clinically helpful information clearly and quickly in order to facilitate appropriate action in terms of optimizing patient care outcomes.d | View Page |
| The May -Hegglin anomaly Illustrated in the upper photograph is a cytoplasmic inclusion somewhat resembling a Doehle body. Note that this inclusion is well defined and there is no evidence of toxic granulation in the cytoplasm. When Doehle-like bodies are identified, May-Hegglin anomaly should be considered in the differential diagnosis even though this entity is rare. The May-Hegglin anomaly is an inherited dominant condition in which large 2 - 5 um, basophilic and pyronophilic inclusions are present in granulocytes, including neutrophils, eosinophils, basophils, and monocytes. Similar to Doehle bodies, the May-Hegglin inclusions also are composed of RNA, probably derived from the rough endoplasmic reticulum. May-Hegglin anomaly includes giant platelets containing few fine granules (lower photograph). Sometimes the platelets have bizarre shapes and variable sizes. Variable degrees of thrombocytopenia complicated by mild bleeding problems and purpura may accompany the aberrant platelets. | View Page |
| Case History A 17-year-old young woman was admitted to the hospital with abdominal pain and a tentative diagnosis of appendicitis.The total white blood count was 14,500 cells/cumm with a left shift and neutrophils with changes tagged by the arrow in the photographs (see blue arrow).The bluish-staining, blurred accumulations in the cytoplasm (Doehle bodies), are located at the cell periphery in neutrophils with toxic changes.Doehle bodies are remnants of endocytoplasmic reticulum and are products of cytokine activity in the induction and shortened activity of neutrophil activation.They are often present in conditions with increased neutrophil lysosomal activity, manifest as toxic granulation.In this case, the presence of Doehle bodies serves as markers for infection-induced leukocytosis and supports the diagnosis of acute appendicitis. | View Page |
| A peripheral blood smear is submitted for morphology review. The patient is a 10 year-old boy with symptoms suggesting appendicitis and an appendectomy is being considered. The total WBC is 18.5 X 1000/uL, RBC's = 5.45 X 1M/uL, hemoglobin = 16.0 g/dL, hematocrit 48.2%;wbc differential: Segs = 53%, bands = 42% (two of which are shown in the photograph), monocytes = 2%, and lymphocytes= 2%. These findings support the diagnosis of appendicitis. | View Page |
| The neutrophils seen in two fields in the upper and lower photographs are representative of a majority of the left shift neutrophils found in this peripheral blood smear. The diagnosis of Pelger-Huet anomaly can be made. | View Page |
| Case Follow-up Illustrated in the upper and lower photographs are two-lobed, eye glass ("pince nez") nuclei of neutrophils typical for patients with Pelger-Huet anomaly. In addition to the characteristic two lobes connected by a delicate bridge, the dense, homogeneous nuclear chromatin helps to define Pelger-Huet anomaly. Since the peripheral blood smear did not support the diagnosis of appendicitis in this patient, and since abdominal pain localized to the right lower quadrant never developed, the boy was hydrated with intravenous fluid and observed. After hydration, his constitutional symptoms improved and the abdominal pain subsided. In fact, the lad was back on the ski slopes the next afternoon. People entering high altitude where the humidity may be very low are susceptible to dehydration and may experience symptoms related to mountain sickness. Therefore, close observation and hydration may be the best practice in monitoring patients with stories and findings similar to this one. A further lesson here is that technologists must be alert to the possibility of Pelger-Huet anomaly if a high white blood cell count with a high percentage of band neutrophils with strikingly uniform morphology and without toxic granulation are found. Inappropriate therapy or an invasive procedure as was contemplated here may be avoided by a proper smear assessment and clinical corroboration. | View Page |
| A most useful follow-up test to consider when faced with hypersegmented neutrophils and oval macrocytes (see photograph) in a peripheral blood smear is: | View Page |
| The cell bulging with inclusions in the image on the right is most consistent with Chediak-Higashi anomaly. | View Page |
| The large blue staining cells represented here in the photographs comprise 50% of the total white blood count.This picture is most consistent with: | View Page |
| Multiple myeloma Plasma cells are uncommonly observed in the peripheral blood smear.They are normal constituents of lymph nodes, spleen, connective tissue and bone marrow. The presence of plasma cells in the peripheral blood is indicative of a large number of conditions mostly related to infections , immune disorders, malignancies, toxic exposures, hypersensitivity reactions and their responses.Although mature plasma cells have a distinct appearance, they still may be confused morphologically with immature plasma cells and other cells with inclusions, reactive changes or nucleated red bloods cell with altered identities.In the upper and lower photographs are plasma cells with features mindful of myeloma cellsThe large myeloma cell in the upper photograph has an eccentric immature nucleus with a muddy chromatin pattern.Note also clumping and stacking of the erythrocytes, bordering on rouleaux formation ,implicating an increase in plasma gamma globulin.The plasma cell with the double nucleus in the lower photograph is particularly suggestive of myeloma.Further studies are in order including a bone marrow examination where at least 30% of bone marrow cells should be variations of mature and immature plasma cells.Serum electrophoresis will reveal a monoclonal globulin spike, and light chains in excess of 1.0 gm/24 hours may be seen in the urine.The presence of lytic bone lesions is a convincing clinical clue.With these findings in combination, a diagnosis of myeloma can be made with assurance. | View Page |