Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR
A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typh...
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| Veröffentlicht in: | Journal of microbiological methods 2017-08, Vol.139, p.150-154 |
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| container_title | Journal of microbiological methods |
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| creator | Prabagaran, Solai Ramatchandirane Kalaiselvi, Vellingiri Chandramouleeswaran, Naganathan Deepthi, Krishnan Nair Geetha Brahmadathan, Kootallur Narayanan Mani, Mariappa |
| description | A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered.
•Alternate method to the Widal test, which is with minimum reliability.•A rapid molecular method with high sensitivity and specificity.•A nested multiplex polymerase chain reaction (PCR) to detect pathogens at low titre. |
| doi_str_mv | 10.1016/j.mimet.2017.05.013 |
| format | Article |
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•Alternate method to the Widal test, which is with minimum reliability.•A rapid molecular method with high sensitivity and specificity.•A nested multiplex polymerase chain reaction (PCR) to detect pathogens at low titre.</description><identifier>ISSN: 0167-7012</identifier><identifier>EISSN: 1872-8359</identifier><identifier>DOI: 10.1016/j.mimet.2017.05.013</identifier><identifier>PMID: 28545919</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Bacterial Proteins - genetics ; Blood - microbiology ; Blood samples ; DNA Primers ; DNA, Bacterial - blood ; Female ; Flagellin - genetics ; Flagellin gene ; Genomic Islands - genetics ; Humans ; Male ; Molecular Diagnostic Techniques - methods ; Multiplex Polymerase Chain Reaction - methods ; Nested multiplex PCR ; Operon ; Salmonella typhi ; Salmonella typhi - genetics ; Salmonella typhi - isolation & purification ; Salmonella typhi - pathogenicity ; Sensitivity and Specificity ; Transcription Factors - genetics ; Typhoid Fever - diagnosis ; Typhoid Fever - microbiology ; Typhoid-Paratyphoid Vaccines ; Vi polysaccharide ; Virulence ; Widal test</subject><ispartof>Journal of microbiological methods, 2017-08, Vol.139, p.150-154</ispartof><rights>2017 Elsevier B.V.</rights><rights>Copyright © 2017 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c359t-57b6d9fe2574aa09d4769239b5d68c8a099f43d219312b3dfba2c8915380b9ad3</citedby><cites>FETCH-LOGICAL-c359t-57b6d9fe2574aa09d4769239b5d68c8a099f43d219312b3dfba2c8915380b9ad3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0167701217301239$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28545919$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Prabagaran, Solai Ramatchandirane</creatorcontrib><creatorcontrib>Kalaiselvi, Vellingiri</creatorcontrib><creatorcontrib>Chandramouleeswaran, Naganathan</creatorcontrib><creatorcontrib>Deepthi, Krishnan Nair Geetha</creatorcontrib><creatorcontrib>Brahmadathan, Kootallur Narayanan</creatorcontrib><creatorcontrib>Mani, Mariappa</creatorcontrib><title>Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR</title><title>Journal of microbiological methods</title><addtitle>J Microbiol Methods</addtitle><description>A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered.
•Alternate method to the Widal test, which is with minimum reliability.•A rapid molecular method with high sensitivity and specificity.•A nested multiplex polymerase chain reaction (PCR) to detect pathogens at low titre.</description><subject>Bacterial Proteins - genetics</subject><subject>Blood - microbiology</subject><subject>Blood samples</subject><subject>DNA Primers</subject><subject>DNA, Bacterial - blood</subject><subject>Female</subject><subject>Flagellin - genetics</subject><subject>Flagellin gene</subject><subject>Genomic Islands - genetics</subject><subject>Humans</subject><subject>Male</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Multiplex Polymerase Chain Reaction - methods</subject><subject>Nested multiplex PCR</subject><subject>Operon</subject><subject>Salmonella typhi</subject><subject>Salmonella typhi - genetics</subject><subject>Salmonella typhi - isolation & purification</subject><subject>Salmonella typhi - pathogenicity</subject><subject>Sensitivity and Specificity</subject><subject>Transcription Factors - genetics</subject><subject>Typhoid Fever - diagnosis</subject><subject>Typhoid Fever - microbiology</subject><subject>Typhoid-Paratyphoid Vaccines</subject><subject>Vi polysaccharide</subject><subject>Virulence</subject><subject>Widal test</subject><issn>0167-7012</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kEtP3DAURq0KVAbaX1Cp8pJNUj_ixF50gUa0RRoEgnZtObYz45ETp3aCOv8eh6EsWVm6PvfxHQC-YFRihOtv-7J3vZ1KgnBTIlYiTD-AFeYNKThl4gSsMtUUDcLkDJyntEcIM1rxj-CMcFYxgcUKHG6Dt3r2KkLj1HYIySUYOviofB8G672C02HcOagGA92U4JOLs7eDttANMM1ptHqy5gUKzsDWh2BgUv3obYLTLoZ5u4ODTQvUz35y-eMfvF8_fAKnnfLJfn59L8CfH9e_17-Kzd3Pm_XVptA5xFSwpq2N6CxhTaUUEqZqakGoaJmpuea5IrqKGoIFxaSlpmsV0VzkpBy1Qhl6AS6Pc8cY_s75ENm7pJdkgw1zklggimvBRZNRekR1DClF28kxul7Fg8RILs7lXr44l4tziZjMznPX19cFc9tb89bzX3IGvh8Bm2M-ORtl0m5RaFzM9qQJ7t0Fz4Phlck</recordid><startdate>201708</startdate><enddate>201708</enddate><creator>Prabagaran, Solai Ramatchandirane</creator><creator>Kalaiselvi, Vellingiri</creator><creator>Chandramouleeswaran, Naganathan</creator><creator>Deepthi, Krishnan Nair Geetha</creator><creator>Brahmadathan, Kootallur Narayanan</creator><creator>Mani, Mariappa</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201708</creationdate><title>Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR</title><author>Prabagaran, Solai Ramatchandirane ; Kalaiselvi, Vellingiri ; Chandramouleeswaran, Naganathan ; Deepthi, Krishnan Nair Geetha ; Brahmadathan, Kootallur Narayanan ; Mani, Mariappa</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c359t-57b6d9fe2574aa09d4769239b5d68c8a099f43d219312b3dfba2c8915380b9ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Bacterial Proteins - genetics</topic><topic>Blood - microbiology</topic><topic>Blood samples</topic><topic>DNA Primers</topic><topic>DNA, Bacterial - blood</topic><topic>Female</topic><topic>Flagellin - genetics</topic><topic>Flagellin gene</topic><topic>Genomic Islands - genetics</topic><topic>Humans</topic><topic>Male</topic><topic>Molecular Diagnostic Techniques - methods</topic><topic>Multiplex Polymerase Chain Reaction - methods</topic><topic>Nested multiplex PCR</topic><topic>Operon</topic><topic>Salmonella typhi</topic><topic>Salmonella typhi - genetics</topic><topic>Salmonella typhi - isolation & purification</topic><topic>Salmonella typhi - pathogenicity</topic><topic>Sensitivity and Specificity</topic><topic>Transcription Factors - genetics</topic><topic>Typhoid Fever - diagnosis</topic><topic>Typhoid Fever - microbiology</topic><topic>Typhoid-Paratyphoid Vaccines</topic><topic>Vi polysaccharide</topic><topic>Virulence</topic><topic>Widal test</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Prabagaran, Solai Ramatchandirane</creatorcontrib><creatorcontrib>Kalaiselvi, Vellingiri</creatorcontrib><creatorcontrib>Chandramouleeswaran, Naganathan</creatorcontrib><creatorcontrib>Deepthi, Krishnan Nair Geetha</creatorcontrib><creatorcontrib>Brahmadathan, Kootallur Narayanan</creatorcontrib><creatorcontrib>Mani, Mariappa</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of microbiological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Prabagaran, Solai Ramatchandirane</au><au>Kalaiselvi, Vellingiri</au><au>Chandramouleeswaran, Naganathan</au><au>Deepthi, Krishnan Nair Geetha</au><au>Brahmadathan, Kootallur Narayanan</au><au>Mani, Mariappa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR</atitle><jtitle>Journal of microbiological methods</jtitle><addtitle>J Microbiol Methods</addtitle><date>2017-08</date><risdate>2017</risdate><volume>139</volume><spage>150</spage><epage>154</epage><pages>150-154</pages><issn>0167-7012</issn><eissn>1872-8359</eissn><abstract>A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered.
•Alternate method to the Widal test, which is with minimum reliability.•A rapid molecular method with high sensitivity and specificity.•A nested multiplex polymerase chain reaction (PCR) to detect pathogens at low titre.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>28545919</pmid><doi>10.1016/j.mimet.2017.05.013</doi><tpages>5</tpages></addata></record> |
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| subjects | Bacterial Proteins - genetics Blood - microbiology Blood samples DNA Primers DNA, Bacterial - blood Female Flagellin - genetics Flagellin gene Genomic Islands - genetics Humans Male Molecular Diagnostic Techniques - methods Multiplex Polymerase Chain Reaction - methods Nested multiplex PCR Operon Salmonella typhi Salmonella typhi - genetics Salmonella typhi - isolation & purification Salmonella typhi - pathogenicity Sensitivity and Specificity Transcription Factors - genetics Typhoid Fever - diagnosis Typhoid Fever - microbiology Typhoid-Paratyphoid Vaccines Vi polysaccharide Virulence Widal test |
| title | Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR |
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