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Mol Cell Probes. 2013 Apr;27(2):86-9. doi: 10.1016/j.mcp.2012.11.003. Epub 2012 Nov 26.

Utilization of a real-time PCR approach for Haemophilus influenzae serotype determination as an alternative to the slide agglutination test.

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  • 1Wadsworth Center, New York State Department of Health, PO Box 22002, Albany, NY 12201-2002, USA.


Our laboratory has developed a simple two-step multiplex real-time PCR for use on isolates of Haemophilus influenzae for molecular serotype identification and the detection of capsular gene targets. The assay consists of a 2-plex real-time PCR targeting the capsule transport gene (bexA), and serotype b specific gene (bcsB), and a 5-plex real-time PCR detecting serotypes a, c, d, e, and f targeting Region II serotype-specific genes. Both real-time PCR assays are highly sensitive (<8 CFU) for all serotypes and 100% specific when tested by a panel of more than 40 bacterial organisms. A retrospective study of 214 isolates received between 1998 and 2011 were tested and compared against the traditional slide agglutination test (SAT) resulting in 100% concordance. We demonstrate that this two-step real-time PCR approach is more sensitive than previously published PCR assays and provides a simple alternative to the SAT. Reliable, rapid and sensitive H. influenzae serotyping is critical for identifying new emerging strains for epidemiological surveillance.

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