Development of polymerase chain reaction-based diagnostic tests for detection of Malsoor virus & adenovirus isolated from Rousettus species of bats in Maharashtra, India
Identifieur interne : 000F17 ( Main/Exploration ); précédent : 000F16; suivant : 000F18Development of polymerase chain reaction-based diagnostic tests for detection of Malsoor virus & adenovirus isolated from Rousettus species of bats in Maharashtra, India
Auteurs : Anita M. Shete ; Pragya Yadav ; Vimal Kumar ; Tushar Nikam ; Kurosh Mehershahi ; Prasad Kokate ; Deepak Patil ; Devendra T. MouryaSource :
- The Indian Journal of Medical Research [ 0971-5916 ] ; 2017.
Abstract
Bats are recognized as important reservoirs for emerging infectious disease and some unknown viral diseases. Two novel viruses, Malsoor virus (family
For rapid and accurate screening of Malsoor virus and adenovirus a nested polymerase chain reaction and TaqMan-based real-time PCR were developed. Highly conserved region of nucleoprotein gene of phleboviruses and polymerase gene sequence from the Indian bat AdV isolate polyprotein gene were selected respectively for diagnostic assay development of Malsoor virus and AdV. Sensitivity and specificity of assays were calculated and optimized assays were used to screen bat samples.
Molecular diagnostic assays were developed for screening of Malsoor virus and AdV and those were found to be specific. Based on the experiments performed with different parameters, nested PCR was found to be more sensitive than real-time PCR; however, for rapid screening, real-time PCR can be used and further nested PCR can be used for final confirmation or in those laboratories where real-time facility/expertise is not existing.
This study reports the development and optimization of nested RT-PCR and a TaqMan-based real-time PCR for Malsoor virus and AdV. The diagnostic assays can be used for rapid detection of these novel viruses to understand their prevalence among bat population.
Url:
DOI: 10.4103/ijmr.IJMR_1447_15
PubMed: 28574020
PubMed Central: 5460580
Affiliations:
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Shete</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Yadav, Pragya" sort="Yadav, Pragya" uniqKey="Yadav P" first="Pragya" last="Yadav">Pragya Yadav</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kumar, Vimal" sort="Kumar, Vimal" uniqKey="Kumar V" first="Vimal" last="Kumar">Vimal Kumar</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Nikam, Tushar" sort="Nikam, Tushar" uniqKey="Nikam T" first="Tushar" last="Nikam">Tushar Nikam</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Mehershahi, Kurosh" sort="Mehershahi, Kurosh" uniqKey="Mehershahi K" first="Kurosh" last="Mehershahi">Kurosh Mehershahi</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kokate, Prasad" sort="Kokate, Prasad" uniqKey="Kokate P" first="Prasad" last="Kokate">Prasad Kokate</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Patil, Deepak" sort="Patil, Deepak" uniqKey="Patil D" first="Deepak" last="Patil">Deepak Patil</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Mourya, Devendra T" sort="Mourya, Devendra T" uniqKey="Mourya D" first="Devendra T." last="Mourya">Devendra T. 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Shete</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Yadav, Pragya" sort="Yadav, Pragya" uniqKey="Yadav P" first="Pragya" last="Yadav">Pragya Yadav</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kumar, Vimal" sort="Kumar, Vimal" uniqKey="Kumar V" first="Vimal" last="Kumar">Vimal Kumar</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Nikam, Tushar" sort="Nikam, Tushar" uniqKey="Nikam T" first="Tushar" last="Nikam">Tushar Nikam</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Mehershahi, Kurosh" sort="Mehershahi, Kurosh" uniqKey="Mehershahi K" first="Kurosh" last="Mehershahi">Kurosh Mehershahi</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kokate, Prasad" sort="Kokate, Prasad" uniqKey="Kokate P" first="Prasad" last="Kokate">Prasad Kokate</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Patil, Deepak" sort="Patil, Deepak" uniqKey="Patil D" first="Deepak" last="Patil">Deepak Patil</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Mourya, Devendra T" sort="Mourya, Devendra T" uniqKey="Mourya D" first="Devendra T." last="Mourya">Devendra T. Mourya</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">The Indian Journal of Medical Research</title><idno type="ISSN">0971-5916</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec id="st1"><title>Background & objectives:</title><p>Bats are recognized as important reservoirs for emerging infectious disease and some unknown viral diseases. Two novel viruses, Malsoor virus (family <italic>Bunyaviridae</italic>, genus, <italic>Phlebovirus</italic>) and a novel adenovirus (AdV) (family, <italic>Adenoviridae</italic> genus, <italic>Mastadenovirus</italic>), were identified from <italic>Rousettus</italic> bats in the Maharashtra State of India. This study was done to develop and optimize real time reverse transcription - polymerase chain reaction (RT-PCR) assays for Malsoor virus and real time and nested PCR for adenovirus from <italic>Rousettus</italic> bats.</p></sec><sec id="st2"><title>Methods:</title><p>For rapid and accurate screening of Malsoor virus and adenovirus a nested polymerase chain reaction and TaqMan-based real-time PCR were developed. Highly conserved region of nucleoprotein gene of phleboviruses and polymerase gene sequence from the Indian bat AdV isolate polyprotein gene were selected respectively for diagnostic assay development of Malsoor virus and AdV. Sensitivity and specificity of assays were calculated and optimized assays were used to screen bat samples.</p></sec><sec id="st3"><title>Results:</title><p>Molecular diagnostic assays were developed for screening of Malsoor virus and AdV and those were found to be specific. Based on the experiments performed with different parameters, nested PCR was found to be more sensitive than real-time PCR; however, for rapid screening, real-time PCR can be used and further nested PCR can be used for final confirmation or in those laboratories where real-time facility/expertise is not existing.</p></sec><sec id="st4"><title>Interpretation & conclusions:</title><p>This study reports the development and optimization of nested RT-PCR and a TaqMan-based real-time PCR for Malsoor virus and AdV. 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Mourya</name><name sortKey="Nikam, Tushar" sort="Nikam, Tushar" uniqKey="Nikam T" first="Tushar" last="Nikam">Tushar Nikam</name><name sortKey="Patil, Deepak" sort="Patil, Deepak" uniqKey="Patil D" first="Deepak" last="Patil">Deepak Patil</name><name sortKey="Shete, Anita M" sort="Shete, Anita M" uniqKey="Shete A" first="Anita M." last="Shete">Anita M. Shete</name><name sortKey="Yadav, Pragya" sort="Yadav, Pragya" uniqKey="Yadav P" first="Pragya" last="Yadav">Pragya Yadav</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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