Identification of Five Novel Salmonella Typhi-Specific Genes as Markers for Diagnosis of Typhoid Fever Using Single-Gene Target PCR Assays
Salmonella Typhi (S. Typhi) causes typhoid fever which is a disease characterised by high mortality and morbidity worldwide. In order to curtail the transmission of this highly infectious disease, identification of new markers that can detect the pathogen is needed for development of sensitive and s...
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| Veröffentlicht in: | BioMed research international 2016-01, Vol.2016 (2016), p.1-9 |
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| creator | Phua, Kia Kien Abdul Rahman, Zaidah Ja’afar, Ja’afar Nuhu Yeoh, Chiann Ying Tan, Clarissa Ling Ling Chin, Kai Ling Goay, Yuan Xin Chinni, Suresh Venkata |
| description | Salmonella Typhi (S. Typhi) causes typhoid fever which is a disease characterised by high mortality and morbidity worldwide. In order to curtail the transmission of this highly infectious disease, identification of new markers that can detect the pathogen is needed for development of sensitive and specific diagnostic tests. In this study, genomic comparison of S. Typhi with other enteric pathogens was performed, and 6 S. Typhi genes, that is, STY0201, STY0307, STY0322, STY0326, STY2020, and STY2021, were found to be specific in silico. Six PCR assays each targeting a unique gene were developed to test the specificity of these genes in vitro. The diagnostic sensitivities and specificities of each assay were determined using 39 S. Typhi, 62 non-Typhi Salmonella, and 10 non-Salmonella clinical isolates. The results showed that 5 of these genes, that is, STY0307, STY0322, STY0326, STY2020, and STY2021, demonstrated 100% sensitivity (39/39) and 100% specificity (0/72). The detection limit of the 5 PCR assays was 32 pg for STY0322, 6.4 pg for STY0326, STY2020, and STY2021, and 1.28 pg for STY0307. In conclusion, 5 PCR assays using STY0307, STY0322, STY0326, STY2020, and STY2021 were developed and found to be highly specific at single-gene target resolution for diagnosis of typhoid fever. |
| doi_str_mv | 10.1155/2016/8905675 |
| format | Article |
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Typhi) causes typhoid fever which is a disease characterised by high mortality and morbidity worldwide. In order to curtail the transmission of this highly infectious disease, identification of new markers that can detect the pathogen is needed for development of sensitive and specific diagnostic tests. In this study, genomic comparison of S. Typhi with other enteric pathogens was performed, and 6 S. Typhi genes, that is, STY0201, STY0307, STY0322, STY0326, STY2020, and STY2021, were found to be specific in silico. Six PCR assays each targeting a unique gene were developed to test the specificity of these genes in vitro. The diagnostic sensitivities and specificities of each assay were determined using 39 S. Typhi, 62 non-Typhi Salmonella, and 10 non-Salmonella clinical isolates. The results showed that 5 of these genes, that is, STY0307, STY0322, STY0326, STY2020, and STY2021, demonstrated 100% sensitivity (39/39) and 100% specificity (0/72). The detection limit of the 5 PCR assays was 32 pg for STY0322, 6.4 pg for STY0326, STY2020, and STY2021, and 1.28 pg for STY0307. In conclusion, 5 PCR assays using STY0307, STY0322, STY0326, STY2020, and STY2021 were developed and found to be highly specific at single-gene target resolution for diagnosis of typhoid fever.</description><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2016/8905675</identifier><identifier>PMID: 27975062</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Publishing Corporation</publisher><subject>Antigens ; Bacteria ; Bacterial genetics ; Biomarkers - metabolism ; Biomedical research ; Comparative analysis ; Deoxyribonucleic acid ; Diagnosis ; Disease transmission ; DNA ; E coli ; Fever ; Genes ; Genes, Bacterial ; Genetic aspects ; Genomes ; Health aspects ; Hospitals ; Laboratories ; Methods ; Mortality ; Parasitology ; Pathogens ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Proteins ; Salmonella ; Salmonella typhi ; Salmonella typhi - genetics ; Sensitivity and Specificity ; Software ; Species Specificity ; Tropical diseases ; Typhoid ; Typhoid fever ; Typhoid Fever - diagnosis ; Typhoid Fever - genetics ; Typhoid Fever - microbiology</subject><ispartof>BioMed research international, 2016-01, Vol.2016 (2016), p.1-9</ispartof><rights>Copyright © 2016 Yuan Xin Goay et al.</rights><rights>COPYRIGHT 2016 John Wiley & Sons, Inc.</rights><rights>Copyright © 2016 Yuan Xin Goay et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2016 Yuan Xin Goay et al. 2016</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c494t-e0fd54f49a83b2e7c4487ea9ad1d6caa9b9d47212df84816bd0c508e60345163</citedby><cites>FETCH-LOGICAL-c494t-e0fd54f49a83b2e7c4487ea9ad1d6caa9b9d47212df84816bd0c508e60345163</cites><orcidid>0000-0002-8626-5848 ; 0000-0001-8999-028X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5126401/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5126401/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27975062$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Chang, Hao-Teng</contributor><creatorcontrib>Phua, Kia Kien</creatorcontrib><creatorcontrib>Abdul Rahman, Zaidah</creatorcontrib><creatorcontrib>Ja’afar, Ja’afar Nuhu</creatorcontrib><creatorcontrib>Yeoh, Chiann Ying</creatorcontrib><creatorcontrib>Tan, Clarissa Ling Ling</creatorcontrib><creatorcontrib>Chin, Kai Ling</creatorcontrib><creatorcontrib>Goay, Yuan Xin</creatorcontrib><creatorcontrib>Chinni, Suresh Venkata</creatorcontrib><title>Identification of Five Novel Salmonella Typhi-Specific Genes as Markers for Diagnosis of Typhoid Fever Using Single-Gene Target PCR Assays</title><title>BioMed research international</title><addtitle>Biomed Res Int</addtitle><description>Salmonella Typhi (S. Typhi) causes typhoid fever which is a disease characterised by high mortality and morbidity worldwide. In order to curtail the transmission of this highly infectious disease, identification of new markers that can detect the pathogen is needed for development of sensitive and specific diagnostic tests. In this study, genomic comparison of S. Typhi with other enteric pathogens was performed, and 6 S. Typhi genes, that is, STY0201, STY0307, STY0322, STY0326, STY2020, and STY2021, were found to be specific in silico. Six PCR assays each targeting a unique gene were developed to test the specificity of these genes in vitro. The diagnostic sensitivities and specificities of each assay were determined using 39 S. Typhi, 62 non-Typhi Salmonella, and 10 non-Salmonella clinical isolates. The results showed that 5 of these genes, that is, STY0307, STY0322, STY0326, STY2020, and STY2021, demonstrated 100% sensitivity (39/39) and 100% specificity (0/72). The detection limit of the 5 PCR assays was 32 pg for STY0322, 6.4 pg for STY0326, STY2020, and STY2021, and 1.28 pg for STY0307. In conclusion, 5 PCR assays using STY0307, STY0322, STY0326, STY2020, and STY2021 were developed and found to be highly specific at single-gene target resolution for diagnosis of typhoid fever.</description><subject>Antigens</subject><subject>Bacteria</subject><subject>Bacterial genetics</subject><subject>Biomarkers - metabolism</subject><subject>Biomedical research</subject><subject>Comparative analysis</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis</subject><subject>Disease transmission</subject><subject>DNA</subject><subject>E coli</subject><subject>Fever</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genetic aspects</subject><subject>Genomes</subject><subject>Health aspects</subject><subject>Hospitals</subject><subject>Laboratories</subject><subject>Methods</subject><subject>Mortality</subject><subject>Parasitology</subject><subject>Pathogens</subject><subject>Polymerase chain reaction</subject><subject>Polymerase Chain Reaction - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BioMed research international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Phua, Kia Kien</au><au>Abdul Rahman, Zaidah</au><au>Ja’afar, Ja’afar Nuhu</au><au>Yeoh, Chiann Ying</au><au>Tan, Clarissa Ling Ling</au><au>Chin, Kai Ling</au><au>Goay, Yuan Xin</au><au>Chinni, Suresh Venkata</au><au>Chang, Hao-Teng</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of Five Novel Salmonella Typhi-Specific Genes as Markers for Diagnosis of Typhoid Fever Using Single-Gene Target PCR Assays</atitle><jtitle>BioMed research international</jtitle><addtitle>Biomed Res Int</addtitle><date>2016-01-01</date><risdate>2016</risdate><volume>2016</volume><issue>2016</issue><spage>1</spage><epage>9</epage><pages>1-9</pages><issn>2314-6133</issn><eissn>2314-6141</eissn><abstract>Salmonella Typhi (S. Typhi) causes typhoid fever which is a disease characterised by high mortality and morbidity worldwide. In order to curtail the transmission of this highly infectious disease, identification of new markers that can detect the pathogen is needed for development of sensitive and specific diagnostic tests. In this study, genomic comparison of S. Typhi with other enteric pathogens was performed, and 6 S. Typhi genes, that is, STY0201, STY0307, STY0322, STY0326, STY2020, and STY2021, were found to be specific in silico. Six PCR assays each targeting a unique gene were developed to test the specificity of these genes in vitro. The diagnostic sensitivities and specificities of each assay were determined using 39 S. Typhi, 62 non-Typhi Salmonella, and 10 non-Salmonella clinical isolates. The results showed that 5 of these genes, that is, STY0307, STY0322, STY0326, STY2020, and STY2021, demonstrated 100% sensitivity (39/39) and 100% specificity (0/72). The detection limit of the 5 PCR assays was 32 pg for STY0322, 6.4 pg for STY0326, STY2020, and STY2021, and 1.28 pg for STY0307. In conclusion, 5 PCR assays using STY0307, STY0322, STY0326, STY2020, and STY2021 were developed and found to be highly specific at single-gene target resolution for diagnosis of typhoid fever.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Publishing Corporation</pub><pmid>27975062</pmid><doi>10.1155/2016/8905675</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-8626-5848</orcidid><orcidid>https://orcid.org/0000-0001-8999-028X</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Antigens Bacteria Bacterial genetics Biomarkers - metabolism Biomedical research Comparative analysis Deoxyribonucleic acid Diagnosis Disease transmission DNA E coli Fever Genes Genes, Bacterial Genetic aspects Genomes Health aspects Hospitals Laboratories Methods Mortality Parasitology Pathogens Polymerase chain reaction Polymerase Chain Reaction - methods Proteins Salmonella Salmonella typhi Salmonella typhi - genetics Sensitivity and Specificity Software Species Specificity Tropical diseases Typhoid Typhoid fever Typhoid Fever - diagnosis Typhoid Fever - genetics Typhoid Fever - microbiology |
| title | Identification of Five Novel Salmonella Typhi-Specific Genes as Markers for Diagnosis of Typhoid Fever Using Single-Gene Target PCR Assays |
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