Reverse transcription polymerase chain reaction‐based method for selectively detecting vegetative cells of toxigenic Clostridium difficile. (November 2014)

• Record Type: Journal Article
• Authors: Senoh, Mitsutoshi; Kato, Haru; Murase, Tomoko; Hagiya, Hideharu; Tagashira, Yasuaki; Fukuda, Tadashi; Iwaki, Masaaki; Yamamoto, Akihiko; Shibayama, Keigo
• Is Part Of: Microbiology and immunology. Volume 58:Number 11(2014:Nov.)
• Page Start: 615
• Page End: 620
• Publication Date: 2014-11
• Subjects: Microbiology -- Periodicals; Immunology -- Periodicals; Allergy and Immunology -- Periodicals; Microbiology -- Periodicals; Microbiologie -- Périodiques; Immunologie -- Périodiques; 579
• DOI: 10.1111/1348-0421.12189
• Languages: English
• ISSNs: 0385-5600

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <sec id="mim12189-sec-0001" sec-type="section"> <p>The laboratory diagnostic methods for <italic>Clostridium difficile</italic> infection (CDI) include toxigenic culture, enzyme immunoassays (EIAs) to detect the toxins of <italic>C. difficile</italic>, and nucleic acid amplification tests (NAATs) to detect <italic>C. difficile</italic> toxin genes, but each of these methods has disadvantages; toxigenic cultures require a long time to produce results, EIAs have low sensitivity, and NAATs that target DNA cannot distinguish vegetative cells from spores and dead cells. Here we report a new detection method that uses reverse transcription polymerase chain reaction to target the toxin‐gene transcripts. This method was able to specifically detect the vegetative cells of toxigenic <italic>C. difficile</italic> in fecal samples in spike tests, with a minimum detection limit of 5 × 10<sup>2</sup> colony‐forming units per 100 mg of stool specimen. The performance of this method was also demonstrated in a pilot scale evaluation using clinical fecal specimens, which showed that this method may be more sensitive than EIA and requires a shorter time than toxigenic culture. This method could potentially be applied in the clinical laboratory to detect <italic>C. difficile</italic> in fecal specimens. The ability of this method to discriminate the presence of vegetative cells from spores and dead cells could help to further the understanding of CDI.</p> </sec> </abstract>