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Modification and evaluation of the Carba NP test by use of paper strip for simple and rapid detection of carbapenemase-producing Enterobacteriaceae

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Abstract

Carbapenemase-producing Enterobacteriaceae (CPE) isolates have now emerged worldwide. We therefore modified the phenotypic Carba NP test by use of a filter paper strip for easily and rapidly identifying CPE in routine laboratory. A collection of 56 CPE and carbapenemase-producing Pseudomonas spp. isolates (including 28 NDM-1, 11 IMP-14a, 1 IMP-1, 1 IMP-4, 1 IMP-9, 1 IMP-15, 4 VIM-2, 1 VIM-1, 1 IMP-14a with VIM-2, 3 OXA-48, 3 OXA-181 and 1 KPC-2 producers) and 41 non-CPE isolates (including 19 ESBL, 7 pAmpC, 3 AmpC, 9 ESBL with pAmpC and 3 non-ESBL & non-AmpC producers) as confirmed by the PCR methods were tested by the paper strip method using pharmaceutical imipenem/cilastatin as a substrate. Bacterial colonies of each isolate were applied directly on filter paper strips dropped with either imipenem-phenol red (test strip) or phenol red solution alone (control strip). The reaction was read within 5 min. This test failed to detect 3 OXA-181, 2 OXA-48 and 3 IMP-14a producers (85.7 % sensitivity), whereas no false positives were seen (100 % specificity). Further evaluation of the paper strip test in 267 CPE screening-positive isolates from three hospitals by their medical technologists showed 92.0 % sensitivity (100 % for NDM producers) and 100 % specificity compared with the PCR methods. Because of its ease, rapidness and cost effective, the paper strip test has a potential for routine CPE testing in low-resource laboratories particularly in areas with high prevalence of NDM enzymes, leading to appropriate antimicrobial therapy and infection control strategy.

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Acknowledgments

This work was supported by Khon Kaen University Research Grant (Grant Number 570029). We thank the CMDL, Khon Kaen University for providing support for Arpasiri Srisrattakarn. We are grateful to Professor David Livermore, Antibiotic Resistance Monitoring & Reference Laboratory, HPA Microbiology Services-Colindale Unit, London, UK; Professor George A Jacoby, Lahey Clinic, Burlington, Massachusetts, USA; Dr. Jianhui Xiong, Clinical Microbiology Fellow at the Department of Laboratory Medicine, University of Toronto, Canada; and DMST Culture Collection, National Institute of Health, Thailand for kindly providing the reference strains and to staff of clinical microbiology laboratories in Srinagarind Hospital, QSNICH and Nan Hospital for collecting the clinical isolates.

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Authors and Affiliations

  1. Medical Sciences Programme, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Arpasiri Srisrattakarn

  2. Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Aroonlug Lulitanond, Chotechana Wilailuckana, Nicha Charoensri & Aroonwadee Chanawong

  3. Clinical Microbiology Unit, Srinagarind Hospital, Khon Kaen University, Khon Kaen, 40002, Thailand

    Lumyai Wonglakorn

  4. Clinical Microbiology Laboratory, The Queen Sirikit National Institute of Child Health, Bangkok, 10400, Thailand

    Sirikan Piyapatthanakul

  5. Clinical Microbiology Laboratory, Nan Hospital, Nan, 55000, Thailand

    Ampai Supajeen

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  1. Arpasiri Srisrattakarn
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  2. Aroonlug Lulitanond
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  3. Chotechana Wilailuckana
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  8. Aroonwadee Chanawong
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Correspondence to Aroonwadee Chanawong.

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Srisrattakarn, A., Lulitanond, A., Wilailuckana, C. et al. Modification and evaluation of the Carba NP test by use of paper strip for simple and rapid detection of carbapenemase-producing Enterobacteriaceae . World J Microbiol Biotechnol 32, 117 (2016). https://doi.org/10.1007/s11274-016-2064-x

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