Evidence map›Paper›PMID 29265030›Full record

ArticleThe Indian journal of medical research2017

A simplified multiplex PCR-based typing method for common

Probodh Borah, Steffen Porwollik, Prerak Desai, Prakash Nayak, Partha Pratim Borah, Pui Cheng, Michael McClelland

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Article in The Indian journal of medical research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

7 authors.

Probodh BorahDepartment of Microbiology, College of Veterinary Science, Assam Agricultural University, Guwahati, India.
Steffen PorwollikDeparment of Microbiology & Molecular Genetics, University of California, Irvine, USA.
Prerak DesaiDeparment of Microbiology & Molecular Genetics, University of California, Irvine, USA.
Prakash NayakDepartment of Bioinformatics Centre, College of Veterinary Science, Assam Agricultural University, Guwahati, India.
Partha Pratim BorahDepartment of Animal Biotechnology, College of Veterinary Science, Assam Agricultural University, Guwahati, India.
Pui ChengDeparment of Microbiology & Molecular Genetics, University of California, Irvine, USA.
Michael McClellandDeparment of Microbiology & Molecular Genetics, University of California, Irvine, USA.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

BACKGROUND &

objectivesA rapid and simple alternative method is needed to replace the laborious, time-consuming Salmonella serotyping. The objective of the present study was to improve and simplify a previously reported multiplex polymerase chain reaction (PCR)-based method and to create an online server to enable rapid determination of serovars.

methodsA method of multiplex PCR-based genome typing (MPGT) was standardized using 59 Salmonella isolates of 31 serovars. Several previously reported primers were modified to obtain a more accurate performance. The screen was separated into four different multiplex reactions distinguishable on standard electrophoresis. A blind study was subsequently performed with 81 isolates of 10 serovars most prevalent in India. Whole genome information from 440 Salmonella isolates was used to confirm the usefulness of this method and concurrence of in silico predictions and PCR results were investigated. A public server (http://www.mpgt-salmonella.res.in) was established for data storage and determination of closest previously observed Salmonella isolates based on obtained MPGT patterns.

resultsThe 16 target genes amplified showed variability in their presence in strains from different serotypes. Hence, identical amplification patterns suggested genetic relatedness of strains and usually identical serological behaviour. The observed absence/presence patterns of genes were converted to an MPGT code. Altogether, 83 different codes were predicted in silico based on the whole genome information of 440 strains. Results confirmed that major serovars usually displayed unique MPGT codes. INTERPRETATION &

conclusionsThe multiplex PCR assay resulted in specific binary codes for isolates from each of the 31 Salmonella serovars tested. The online server allowed the user to compare obtained PCR results with stored previous patterns. Simplicity, speed and cost-effectiveness make this tool useful for quick outbreak management.

Indexed as

Bacterial Typing TechniquesHumansIndiaMultiplex Polymerase Chain ReactionSalmonella entericaSalmonella InfectionsSerogroupSerotyping

Identifiers

PMID29265030
PMCPMC5761039

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