Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase

Masanori Watahiki; Ryuji Kawahara; Masahiro Suzuki; Miyako Aoki; Kaoru Uchida; Yuko Matsumoto; Yuko Kumagai; Makiko Noda; Kanako Masuda; Chiemi Fukuda; Seiya Harada; Keiko Senba; Masato Suzuki; Mari Matsui; Satowa Suzuki; Keigo Shibayama; Hiroto Shinomiya

doi:10.7883/yoken.JJID.2019.041

Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase

Masanori Watahiki, Ryuji Kawahara, Masahiro Suzuki, Miyako Aoki, Kaoru Uchida, Yuko Matsumoto, Yuko Kumagai, Makiko Noda, Kanako Masuda, Chiemi Fukuda, Seiya Harada, Keiko Senba, Masato Suzuki, Mari Matsui, Satowa Suzuki, Keigo Shibayama, Hiroto Shinomiya

Research output: Contribution to journal › Article › peer-review

18 Citations (Scopus)

Abstract

A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: bla bla bla bla bla and bla Of 70 bla_IMP variants, 67 subtypes were simulated to^KPC,be PCR-positive^IMP,^NDM,based^VIM,on in silico^OXA-48-like,simulation and^GES.the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.

Original language	English
Pages (from-to)	166-172
Number of pages	7
Journal	Japanese journal of infectious diseases
Volume	73
Issue number	2
DOIs	https://doi.org/10.7883/yoken.JJID.2019.041
Publication status	Published - 2020
Externally published	Yes

All Science Journal Classification (ASJC) codes

Microbiology (medical)
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.7883/yoken.JJID.2019.041

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Watahiki, M., Kawahara, R., Suzuki, M., Aoki, M., Uchida, K., Matsumoto, Y., Kumagai, Y., Noda, M., Masuda, K., Fukuda, C., Harada, S., Senba, K., Suzuki, M., Matsui, M., Suzuki, S., Shibayama, K., & Shinomiya, H. (2020). Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase. Japanese journal of infectious diseases, 73(2), 166-172. https://doi.org/10.7883/yoken.JJID.2019.041

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title = "Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase",

abstract = "A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: bla bla bla bla bla and bla Of 70 blaIMP variants, 67 subtypes were simulated toKPC,be PCR-positiveIMP,NDM,basedVIM,on in silicoOXA-48-like,simulation andGES.the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.",

author = "Masanori Watahiki and Ryuji Kawahara and Masahiro Suzuki and Miyako Aoki and Kaoru Uchida and Yuko Matsumoto and Yuko Kumagai and Makiko Noda and Kanako Masuda and Chiemi Fukuda and Seiya Harada and Keiko Senba and Masato Suzuki and Mari Matsui and Satowa Suzuki and Keigo Shibayama and Hiroto Shinomiya",

year = "2020",

doi = "10.7883/yoken.JJID.2019.041",

language = "English",

volume = "73",

pages = "166--172",

journal = "Japanese journal of infectious diseases",

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Watahiki, M, Kawahara, R, Suzuki, M, Aoki, M, Uchida, K, Matsumoto, Y, Kumagai, Y, Noda, M, Masuda, K, Fukuda, C, Harada, S, Senba, K, Suzuki, M, Matsui, M, Suzuki, S, Shibayama, K & Shinomiya, H 2020, 'Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase', Japanese journal of infectious diseases, vol. 73, no. 2, pp. 166-172. https://doi.org/10.7883/yoken.JJID.2019.041

TY - JOUR

T1 - Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase

AU - Watahiki, Masanori

AU - Kawahara, Ryuji

AU - Suzuki, Masahiro

AU - Aoki, Miyako

AU - Uchida, Kaoru

AU - Matsumoto, Yuko

AU - Kumagai, Yuko

AU - Noda, Makiko

AU - Masuda, Kanako

AU - Fukuda, Chiemi

AU - Harada, Seiya

AU - Senba, Keiko

AU - Suzuki, Masato

AU - Matsui, Mari

AU - Suzuki, Satowa

AU - Shibayama, Keigo

AU - Shinomiya, Hiroto

PY - 2020

Y1 - 2020

N2 - A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: bla bla bla bla bla and bla Of 70 blaIMP variants, 67 subtypes were simulated toKPC,be PCR-positiveIMP,NDM,basedVIM,on in silicoOXA-48-like,simulation andGES.the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.

AB - A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: bla bla bla bla bla and bla Of 70 blaIMP variants, 67 subtypes were simulated toKPC,be PCR-positiveIMP,NDM,basedVIM,on in silicoOXA-48-like,simulation andGES.the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.

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SN - 1344-6304

VL - 73

SP - 166

EP - 172

JO - Japanese journal of infectious diseases

JF - Japanese journal of infectious diseases

IS - 2

ER -

Single-tube multiplex polymerase chain reaction for the detection of genes encoding enterobacteriaceae carbapenemase

Abstract

All Science Journal Classification (ASJC) codes

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