Clinical evaluation of a non-purified direct molecular assay for the detection of Clostridioides difficile toxin genes in stool specimens

Toshinori Hara; Hiromichi Suzuki; Tadatomo Oyanagi; Norito Koyanagi; Akihito Ushiki; Naoki Kawabata; Miki Goto; Yukio Hida; Yuji Yaguchi; Kiyoko Tamai; Shigeyuki Notake; Yosuke Kawashima; Akio Sugiyama; Keiichi Uemura; Seiya Kashiyama; Toru Nanmoku; Satoshi Suzuki; Hiroshi Yamazaki; Hideki Kimura; Hiroyuki Kunishima; Hiroki Ohge

doi:10.1371/journal.pone.0234119

Clinical evaluation of a non-purified direct molecular assay for the detection of Clostridioides difficile toxin genes in stool specimens

PLoS One. 2020 Jun 3;15(6):e0234119. doi: 10.1371/journal.pone.0234119. eCollection 2020.

Authors

Toshinori Hara^{1

2}, Hiromichi Suzuki³, Tadatomo Oyanagi⁴, Norito Koyanagi⁵, Akihito Ushiki⁶, Naoki Kawabata⁷, Miki Goto⁸, Yukio Hida⁹, Yuji Yaguchi¹⁰, Kiyoko Tamai¹⁰, Shigeyuki Notake¹¹, Yosuke Kawashima¹², Akio Sugiyama¹², Keiichi Uemura⁵, Seiya Kashiyama^{1

2}, Toru Nanmoku⁸, Satoshi Suzuki¹³, Hiroshi Yamazaki⁷, Hideki Kimura⁹, Hiroyuki Kunishima¹⁴, Hiroki Ohge¹⁵

Affiliations

¹ Section of Clinical Laboratory, Department of Clinical Practice and Support, Hiroshima University Hospital, Hiroshima, Japan.
² Division of Clinical Laboratory Medicine, Hiroshima University Hospital, Hiroshima, Japan.
³ Division of Infectious Diseases, Department of Medicine, Tsukuba Medical Center Hospital, Ibaraki, Japan.
⁴ Department of Clinical Laboratory, St. Marianna University School of Medicine Hospital, Kanagawa, Japan.
⁵ Department of Clinical Laboratory, Chutoen General Medical Center, Shizuoka, Japan.
⁶ Department of Clinical Laboratory, Tone Chuo Hospital, Gunma, Japan.
⁷ Department of Clinical Laboratory, Tsuruga Municipal Hospital, Fukui, Japan.
⁸ Department of Clinical Laboratory, University of Tsukuba Hospital, Ibaraki, Japan.
⁹ Department of Clinical Laboratory, University of Fukui Hospital, Fukui, Japan.
¹⁰ Miroku Medical Laboratory Inc., Nagano, Japan.
¹¹ Department of Clinical Laboratory, Tsukuba Medical Center Hospital, Ibaraki, Japan.
¹² Diagnostic System Department, TOYOBO Co., Ltd., Osaka, Japan.
¹³ Division of General Medicine, Tone Chuo Hospital, Gunma, Japan.
¹⁴ Department of Infectious Diseases, St. Marianna University School of Medicine, Kanagawa, Japan.
¹⁵ Department of Infectious diseases, Hiroshima University Hospital, Hiroshima, Japan.

Abstract

Recently, a new rapid assay for the detection of tcdB gene of Clostridioides difficile was developed using the GENECUBE. The assay can directly detect the tcdB gene from stool samples without a purification in approximately 35 minutes with a few minutes of preparation process. We performed a prospective comparative study of the performance of the assay at eight institutions in Japan. Fresh residual stool samples (Bristol stool scale ≥5) were used and comparisons were performed with the BD MAX Cdiff assay and toxigenic cultures. For the evaluation of 383 stool samples compared with the BD MAX Cdiff assay, the sensitivity, and specificity of the two assays was 99.0% (379/383), 98.1% (52/53), 99.1% (327/330), respectively. In the comparison with toxigenic culture, the total, sensitivity, and specificity were 96.6% (370/383), 85.0% (51/60), and 98.8% (319/323), respectively. The current investigation indicated the GENECUBE Clostridioides difficile assay has equivalent performance with the BD MAX Cdiff assay for the detection of tcdB gene of C. difficile.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics*
Bacterial Toxins / genetics*
Clostridioides difficile / genetics*
Clostridioides difficile / isolation & purification
Clostridium Infections / diagnosis
DNA, Bacterial / genetics
DNA, Bacterial / metabolism
Enterotoxins / genetics
Feces / microbiology*
Humans
Polymerase Chain Reaction / methods
Reagent Kits, Diagnostic
Repressor Proteins / genetics
Sensitivity and Specificity

Substances

Bacterial Proteins
Bacterial Toxins
DNA, Bacterial
Enterotoxins
Reagent Kits, Diagnostic
Repressor Proteins
TcdC protein, Clostridium difficile
tcdA protein, Clostridium difficile
toxB protein, Clostridium difficile

Grants and funding

This study was supported by TOYOBO Co., Ltd. The funder provided support in the form of salaries to authors A. Sugiyama and Y. Kawashima, and lecture fees to authors H. Suzuki, and advisory fees to author H. Suzuki. The funder did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the Author contributions’ section.