A novel ready-to-use dry-reagent polymerase chain reaction for detection of Escherichia coli & Shigella species

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dc.contributor.authorMishra, MNen_US
dc.contributor.authorKulkarni, RDen_US
dc.contributor.authorMohanraj, Jen_US
dc.contributor.authorNisshanthini, S Den_US
dc.contributor.authorAjantha, G Sen_US
dc.contributor.authorChandrasekhar, Aen_US
dc.contributor.authorKenge, Pen_US
dc.contributor.authorBhat, Sen_US
dc.date.accessioned2020-04-10T01:42:37Z
dc.date.available2020-04-10T01:42:37Z
dc.date.issued2019-05
dc.description.abstractBackground & objectives: Polymerase chain reaction (PCR) has wide acceptance for rapid identification of pathogens and also for diagnosis of infectious conditions. However, because of economic and expertise constraints, a majority of small or peripheral laboratories do not use PCR. The objective of the present study was to develop a dry-reagent PCR assay as an alternative to conventional PCR to assess its applicability in routine laboratory practice using malB gene for identification of Escherichia coli as a model. Methods: A total of 184 isolates were selected for the study comprising clinical isolates of E. coli and non-E. coli including Shigella sp. and a few other control strains. The DNA was isolated from all the isolates. The isolated DNA as well as the overnight grown bacterial cultures were subjected to both conventional wet PCR and dry-reagent PCR. Results: The genomic DNA isolated from E. coli showed amplification of malB gene in both conventional wet and dry-reagent PCR and the band was observed at 491 bp. In dry-reagent PCR, the overnight grown E. coli cells also showed positive result. The non-E. coli strains other than Shigella sp. showed negative in both conventional wet and dry-reagent PCR. Shigella sp. showed positive in both conventional wet and dry-reagent PCR. Interpretation & conclusions: Considering the elimination of genomic DNA isolation step, and similar results with the conventional wet PCR, dry-reagent PCR may be a good alternative for the conventional wet PCR.en_US
dc.identifier.affiliationsDepartment of Molecular Diagnostics, Bhat Biotech India Pvt. Ltd., Bengaluru, Indiaen_US
dc.identifier.affiliationsDepartment of Microbiology, SDM College of Medical Sciences & Hospital, Dharwad, Indiaen_US
dc.identifier.citationMishra MN, Kulkarni RD, Mohanraj J, Nisshanthini S D, Ajantha G S, Chandrasekhar A, Kenge P, Bhat S. A novel ready-to-use dry-reagent polymerase chain reaction for detection of Escherichia coli & Shigella species. Indian Journal of Medical Research. 2019 May; 149(5): 671-676en_US
dc.identifier.issn0971-5916
dc.identifier.issn0975-9174
dc.identifier.placeIndiaen_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/195877
dc.languageenen_US
dc.publisherIndian Council of Medical Researchen_US
dc.relation.issuenumber5en_US
dc.relation.volume149en_US
dc.source.urihttps://dx.doi.org/10.4103/ijmr.IJMR_1394_17en_US
dc.subjectABC transporteren_US
dc.subjectdry-reagent PCRen_US
dc.subjectEscherichia coli- malB geneen_US
dc.subjectmaltose/maltodextrin transporter ATP-binding geneen_US
dc.subjectpathogensen_US
dc.subjectShigella spen_US
dc.titleA novel ready-to-use dry-reagent polymerase chain reaction for detection of Escherichia coli & Shigella speciesen_US
dc.typeJournal Articleen_US
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