191. A Simple and Rapid Detection System for Direct Identification of Carbapenemase-Producing Enterobacteriaceae in Clinical Samples
Session: Poster Abstract Session: Diagnostics: Bacteriology, Sequencing, and Resistance
Thursday, October 27, 2016
Room: Poster Hall
  • Poster IDweek.pdf (969.4 kB)
  • Background:

    Carbapenemase-producing Enterobacteriaceae(CPE) is a global health issue due to their hasty dissemination through the transfer of carbapenemase genes. Hence, rapid detection is necessary to take relevant control measures against CPE infections/colonization. We established a rapid and multiplex CPE detection system - Single Tag Hybridization Printed Array Strip (STH-PAS) by targeting the four different major carbapenemases. STH-PAS is a DNA-DNA hybridization technique where the oligonucleotide tag in the primer of PCR product hybridizes to its probe imprinted on a chromatographic strip without denaturation. Further, the efficacy of STH-PAS in detecting CPE directly in clinical samples is evaluated.


    STH-PAS was tailored to detect various alleles of the four carbapenemase genes – NDM, KPC, IMP, and OXA-48 like in a single reaction. Then, the efficiency of hybridization in STH-PAS for detection of carbapenemases was compared with conventional PCR. The efficiency of carbapenemase detection by STH-PAS was analysed in CPE (n=49) and non-CPE strains (n=10). A total of 114 CPE suspected stool samples were subjected to STH-PAS to examine its utilization for direct clinical samples.


    The ideal conditions for hybridization without non-specificity in STH-PAS was determined. STH-PAS was found to be 10 times more sensitive than conventional PCR techniques. It showed both sensitivity and specificity of 100% for carbapenemase detection in bacterial strains (n=59). As it is not affected by any of the inhibitory substances in clinical specimens, STH-PAS showed 90.91% sensitivity and 98.57% specificity in detecting carbapenemase directly in stool samples (n=114).


    The results of the current study show that STH-PAS possesses several advantages as a good detection system for CPE. As it is very rapid and simple to interpret the results with naked eye, STH-PAS could be applied in poorly resourced countries. It has been planned to assess the effectiveness of STH-PAS as a surveillance tool in clinical settings to control the transmission of CPE.

    Rathina Kumar Shanmuga Kani, M.Tech.1,2, Yukihiro Akeda, PhD.1,2, Norihisa Yamamoto, MD.1,2, Noriko Sakamoto, M.Sc.2, Hideharu Hagiya, MD., PhD.1,2, Hisao Yoshida, MD.1, Takuya Kodera, PhD.3, Pitak Santanirand, PhD.4, Shigeyuki Hamada, DDS., PhD.2 and Kazunori Tomono, MD5, (1)Division of Infection Control and Prevention, Graduate School of Medicine, Osaka University, Suita, Japan, (2)Research Institute for Microbial Diseases, Osaka University, Suita, Japan, (3)Tohoku Bio-Array Co. Ltd.,, Sendai, Japan, (4)Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand, (5)Division of Infection Control and Prevention, Osaka University, Suita, Japan


    R. K. Shanmuga Kani, None

    Y. Akeda, None

    N. Yamamoto, None

    N. Sakamoto, None

    H. Hagiya, None

    H. Yoshida, None

    T. Kodera, None

    P. Santanirand, None

    S. Hamada, None

    K. Tomono, None

    Findings in the abstracts are embargoed until 12:01 a.m. CDT, Wednesday Oct. 26th with the exception of research findings presented at the IDWeek press conferences.