Quantitative real-time PCR assay for Clostridium septicum in poultry gangrenous dermatitis associated samples.

Abstract:

:Clostridium septicum is a spore-forming anaerobe frequently implicated in cases of gangrenous dermatitis (GD) and other spontaneously occurring myonecrotic infections of poultry. Although C. septicum is readily cultured from diseased tissues it can be difficult to enumerate due to its tendency to swarm over the surface of agar plates. In this study a quantitative real-time PCR assay was developed in order to more accurately measure the levels of C. septicum in healthy as well as GD associated poultry samples. The assay was specifically designed to target the C. septicum alpha toxin gene, csa, which is, to our knowledge, carried by all strains of C. septicum and has been shown to be essential for virulence. Genomic DNAs from a diverse collection of bacterial species, including closely related Clostridium chauvoei, Clostridium carnis, Clostridium tertium as well as several strains of Clostridium perfringens, all failed to produce a positive reaction. An approximate reproducible limit of detection in spiked extracts of at least 10(3) cfu/g of C. septicum was observed for a variety of different sample types. C. septicum levels in broiler chicken field samples estimated from the results of qPCR were statistically correlated to culture based enumerations obtained from those same tissues.

journal_name

Mol Cell Probes

authors

Neumann AP,Dunham SM,Rehberger TG,Siragusa GR

doi

10.1016/j.mcp.2010.04.001

subject

Has Abstract

pub_date

2010-08-01 00:00:00

pages

211-8

issue

4

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(10)00021-6

journal_volume

24

pub_type

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