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Gram-positive Information and Courses from MediaLab, Inc.

These are the MediaLab courses that cover Gram-positive and links to relevant pages within the course.

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Chemical Screening of Urine by Reagent Strip
Test Sensitivity

This test is sensitive to 0.06-0.1 mg/dL nitrite ion in urines with a low specific gravity and ascorbic acid concentrations of less than 25 mg/dL. Pink spots or pink edges should not be interpreted as a positive result because some medications can color urine red or turn red in an acid environment. Any degree of uniform pink color should be considered positive, suggesting the presence of 105 organisms/mL. Detection of low levels of nitrite ion may be enhanced by comparing the activated test strip to a white background. It is important to note that color development is NOT proportional to the number of bacteria present. The test is specific for nitrites and does not react with any other substances normally present in urine. Negative results do not necessarily rule out a urinary tract infection because yeasts or gram-positive bacteria unable to reduce nitrites may be the causative agent.

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Nitrates in urine are reduce to nitrites by:View Page

CLIA Microbiology / Serology Review
Which of the following best describes the organisms seen in this illustration:View Page
Which of the following best describes the organisms seen in this illustration:View Page

Current Topics in Clinical Microbiology
The Gram stain report to be issued based on the microscopic characteristics seen in the accompanying picture would most correctly be, "many wbc with..."View Page
Enterococcus ID

Illustrated in this composite photograph is the gram stain of Enterococcus species (upper), illustrating gram positive cocci arranged in pairs and short chains.The viridans streptococci generally form gram positive cocci in long chains; the gram-positive cells of S. pneumoniae, on the other hand, may appear similar in gram stain.In the lower right is shown a 6.5% NaCl tube. The turbidity indicates growth, a unique characteristic for Enterococcus species.In the lower left frame is a picture of a bile esculin hydrolysis slant, showing a positive reaction (black pigment) characteristic of Enterococcus species. The streptococci are uniformly negative.Some strains of Leuconostoc species and Pediococcus species can grow in 6.5% NaCl and are bile-esculin positive; however, they should be ruled out because of the negative PYR reaction.

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Gas gangrene may be seen in infections with all the following clostridia except:View Page
The gram stain shown in the photograph was prepared from a positive anaerobic blood culture bottle after 36 hours incubation. Based on the morphology of the bacterial cells (some with spores--blue arrows), the most likely identification is:View Page
A gram stain of the serous exudate is shown in the photograph. The appropriate report would read:View Page
The Gram stain prepared from the positive blood culture is shown in the photograph. The appropriate report is:View Page
Review 2

Cunningham MW.: Pathogenesis of group A streptococcal infections. Clinical Microbiology Reviews. 13):470-511, 2000Group A streptococci are model extracellular gram-positive pathogens responsible for pharyngitis, impetigo, rheumatic fever, and acute glomerulonephritis. A resurgence of invasive streptococcal diseases and rheumatic fever has appeared in outbreaks over the past 10 years, with a predominant M1 serotype as well as others identified with the outbreaks.Emm (M protein) gene sequencing has changed serotyping, and new virulence genes and new virulence regulatory networks have been defined. The emm gene superfamily has expanded to include antiphagocytic molecules and immunoglobulin-binding proteins with common structural features.At least nine superantigens have been characterized, all of which may contribute to toxic streptococcal syndrome. An emerging theme is the dichotomy between skin and throat strains in their epidemiology and genetic makeup. Eleven adhesions have been reported, and surface plasmin-binding proteins have been defined.The strong resistance of the group A streptococcus to phagocytosis is related to factor H and fibrinogen binding by M protein and to disarming complement component C5a by the C5a peptidase. Molecular mimicry appears to play a role in autoimmune mechanisms involved in rheumatic fever, while nephritis strain-associated proteins may lead to immune-mediated acute glomerulonephritis. Vaccine strategies have focused on recombinant M protein and C5a peptidase vaccines, and mucosal vaccine delivery systems are under investigation.

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A Brown and Brenn gram stain was performed on one of the tissue biopsy specimens. Organisms were seen as shown in the photograph. Based on the history and the appearance of the bacteria, the most likely identification is:View Page
Review 3

Rouquette C. Berche P. The pathogenesis of infection by Listeria monocytogenes Microbiologia. 12:245-58, 1996 Listeria monocytogenes is a Gram-positive bacterium responsible for severe infections in human and a large variety of animal species. It is a facultative intracellular pathogen which invades macrophages and most tissue cells of infected hosts where it can proliferate. The molecular basis of this intracellular parasitism has been to a large extent elucidated. The virulence factors, including internalin, listeriolysin O, phospholipases and a bacterial surface protein, ActA, are encoded by chromosomal genes organized in operons. Following internalisation into host cells, the bacteria escape from the phagosomal compartment and enter the cytoplasm. They then spread from cell to cell by a process involving actin polymerisation. In infected hosts, the bacteria cross the intestinal wall at Peyer's patches to invade the mesenteric lymph nodes and the blood. The main target organ is the liver, where the bacteria multiply inside hepatocytes. Early recruitment of polymorphonuclear cells lead to hepatocyte lysis, and thereby bacterial release This causes prolonged septicaemia, particularly in immunocompromised hosts, thus exposing the placenta and brain to infection. The prognosis of listeriosis depends on the severity of meningoencephalitis, due to the elective location of foci of infection in the brain stem (rhombencephalitis). Despite bactericidal antibiotic therapy, the overall mortality is still high (25 to 30%).

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White Cell and Platelet Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
The inclusions noted in the cytoplasm of this band neutrophil are most suggestive of:View Page


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