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Genetic tests for...

A/B/AB blood type in macaques

Fetal sexing

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Mamu-7 in macaques

CYP2C76 c.449TG>A
in macaques

Mu opioid receptor
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in sooty mangabeys

...and more - contact Zoologix with your genetic testing requirements


Klebsiella PCR test for primates

primate assay data sheet

Klebsiella pneumoniae (including virulence factors magA and rmpA)

Test code:
B0060 - Qualitative ultrasensitive detection of Klebsiella pneumoniae bacteria by real time polymerase chain reaction.  This assay detects and differentiates a genomic target and two virulence factors, magA and rmpA.

Klebsiella pneumoniae bacteria belong to the family Enterobacteriaceae. These organisms are nonmotile, rod-shaped, gram-negative bacteria with a prominent polysaccharide capsule. This capsule encases the entire cell surface and gives the bacterium a large appearance on gram stain; it also imparts resistance against many host defense mechanisms.

Members of the Klebsiella genus typically express 2 types of antigens on their cell surface. One is a lipopolysaccharide (O antigen); the other is a capsular polysaccharide (K antigen). Both of these antigens contribute to pathogenicity of the bacteria in this genus. Approximately 9 O antigens and 77 K antigens exist. The structural variability of these antigens forms the basis for classification of the bacteria into various serotypes. Nevertheless, there are no specific correlations of the various serotypes with pathogenicity.

Besides Klebsiella pneumoniae, the Klebsiella genus also contains Klebsiella ozaenae, Klebsiella rhinoscleromatis, Klebsiella oxytoca, Klebsiella planticola, Klebsiella terrigena, and Klebsiella ornithinolytica. All these bacteria have very similar DNA homology. K pneumoniae is the most medically important species of the group, but K. oxytoca and K. rhinoscleromatis have also been found in human clinical specimens.

Klebsiellae can be found in the environment and have widespread occurrence. In humans, they may colonize the skin, pharynx or gastrointestinal tract. They may also colonize sterile wounds and urine. Many animals and humans are asymptomatic carriers. Klebsiellae may even be regarded as normal flora in parts of the colon, intestinal tract and biliary tract. Oropharyngeal carriage has been associated with endotracheal intubation, impaired host defenses, and antimicrobial use.

On culture, K. pneumoniae produces large, sticky colonies on an agar plate. Some strains of K. pneumoniae possess a “hypermucoviscosity” phenotype; this phenotype is related to higher pathogenicity. The expression of this hypermucoviscosity phenotype is determined by the presence of several genes, including magA (mucoviscosity-associated gene A) and rmpA (regulator of the mucoid phenotype A). The hypermucoviscosity phenotype of K. pneumoniae can be classified as capsular serotypes K1 or K2. K1 serotypes of K. pneumoniae have rmpA and magA. K2 serotypes of K. pneumoniae have rmpA but not magA. Studies have shown that while magA presents exclusively in the K1 capsular serotype, rmpA can be detected in either K1 or K2 capsular serotypes and also in approximately two-thirds of non-K1/K2 capsular serotypes. Capsular serotypes K1 and K2 are reported to play an important role in the invasive ability of K. pneumoniae (Yeh et al., 2007). In some studies, the presence of magA is linked to liver abscess formation (Fang et al., 2004).

Culture detection of K. pneumoniae is of limited value because the method cannot reliably differentiate pathogenic strains from nonpathogenic Klebsiella bacteria present as normal flora in the body. Although hypermucoviscosity can be assessed by a “string test,” the test is highly subjective and can lead to false results. Furthermore, culture identification cannot definitively identify the presence of magA and rmpA which are important in determining pathogenic potential of the bacteria. Molecular detection not only provides rapid, sensitive and specific identification of the bacteria, but can also accurately determine the presence of these virulence factors.

Utilities:

  • Help confirm the disease causing agent
  • Shorten the time required to confirm a clinical diagnosis of Klebsiella pneumoniae infection
  • Help ensure colonies are free of Klebsiella pneumoniae
  • Differentiate virulent from non-virulent Klebsiella pneumoniae strains
  • Identify magA and rmpA virulence factors in Klebsiella pneumoniae strains
  • Early prevention of spread of these bacteria among a facility
  • Minimize human exposure to these bacteria
  • Safety monitoring of biological products and vaccines that derive from primates

References:
Fang, C.T., Chuang, Y.P., Shun, C.T., Chang, S.C. and Wang, J.T. (2004) A novel virulence gene in Klebsiella pneumoniae strains causing primary liver abscess and septic metastatic complications. J. Exp. Med. 199:697–705.
Yeh, K.M., Kurup, A., Siu, L.K., Koh, Y.L., Fung, C.P., Lin, J.C., Chen, T.L., Chang, F.Y. and Koh, T.H.(2007) Capsular serotype K1 or K2, rather than magA and rmpA, is a major virulence determinant for Klebsiella pneumoniae liver abscess in Singapore and Taiwan. J Clin Microbiol 45:466–471.

Specimen requirement:

Preferred specimens - Rectal swab, or oral swab, or 0.2 ml feces, or 0.2 ml bacterial culture.

Less preferred specimen - 0.2 ml whole blood in EDTA (purple top) or ACD (yellow top) tube.

For specimen types other than those listed here, please call to confirm specimen acceptability and shipping instructions.

For all specimen types, if there will be a delay in shipping, or during very warm weather, refrigerate specimens until shipped and ship with a cold pack unless more stringent shipping requirements are specified. Frozen specimens should be shipped so as to remain frozen in transit. See shipping instructions for more information.

Turnaround time: 3 business days

Methodology: Qualitative real time PCR

Normal range: Nondetected

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