Duplex TaqMan Hydrolysis Probe-Based Molecular Assay for Simultaneous Detection and Differentiation of Burkholderia pseudomallei and Leptospira spp. DNA
Melioidosis and leptospirosis, caused by two different bacteria, Burkholderia pseudomallei and Leptospira spp., are potentially fatal infections that share a very similar spectrum of clinical features and cause significant mortality and morbidity in humans and livestock. Early detection is important...
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| Veröffentlicht in: | BioMed research international 2019-01, Vol.2019 (2019), p.1-6 |
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| creator | Ismail, Nabilah Yean Yean, Chan Harun, Azian Aziah, Ismail Mustaffa, Khairul Mohd Fadzli Ismail, Asmaliza S. Goay, Yuan Xin Foo, Phiaw Chong Lih Huey, Lee Mohd Ali, Mohammad Ridhuan Phua, Kia Kien |
| description | Melioidosis and leptospirosis, caused by two different bacteria, Burkholderia pseudomallei and Leptospira spp., are potentially fatal infections that share a very similar spectrum of clinical features and cause significant mortality and morbidity in humans and livestock. Early detection is important for better clinical consequences. To our knowledge, there is no diagnostic tool available to simultaneously detect and differentiate melioidosis and leptospirosis in humans and animals. In this study, we described a duplex TaqMan probe-based qPCR for the detection of B. pseudomallei and Leptospira spp. DNA. The performance of the assay was evaluated on 20 B. pseudomallei isolates, 23 Leptospira strains, and 39 other microorganisms, as well as two sets of serially diluted reference strains. The duplex qPCR assay was able to detect 0.02 pg (~ 4 copies) Leptospira spp. DNA and 0.2 pg (~ 25.6 copies) B. pseudomallei DNA. No undesired amplification was observed in other microorganisms. In conclusion, the duplex qPCR assay was sensitive and specific for the detection of B. pseudomallei & Leptospira spp. DNA and is suitable for further analytical and clinical evaluation. |
| doi_str_mv | 10.1155/2019/9451791 |
| format | Article |
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Early detection is important for better clinical consequences. To our knowledge, there is no diagnostic tool available to simultaneously detect and differentiate melioidosis and leptospirosis in humans and animals. In this study, we described a duplex TaqMan probe-based qPCR for the detection of B. pseudomallei and Leptospira spp. DNA. The performance of the assay was evaluated on 20 B. pseudomallei isolates, 23 Leptospira strains, and 39 other microorganisms, as well as two sets of serially diluted reference strains. The duplex qPCR assay was able to detect 0.02 pg (~ 4 copies) Leptospira spp. DNA and 0.2 pg (~ 25.6 copies) B. pseudomallei DNA. No undesired amplification was observed in other microorganisms. In conclusion, the duplex qPCR assay was sensitive and specific for the detection of B. pseudomallei & Leptospira spp. DNA and is suitable for further analytical and clinical evaluation.</description><identifier>ISSN: 2314-6133</identifier><identifier>ISSN: 2314-6141</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2019/9451791</identifier><identifier>PMID: 31355287</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Publishing Corporation</publisher><subject>Animals ; Assaying ; Burkholderia pseudomallei ; Burkholderia pseudomallei - genetics ; Deoxyribonucleic acid ; Diagnostic software ; Diagnostic systems ; DNA ; DNA, Bacterial - genetics ; Genetic research ; Gram-negative bacteria ; Health aspects ; Humans ; Hydrolysis ; Infection ; Infectious diseases ; Leptospira ; Leptospira - genetics ; Leptospirosis ; Leptospirosis - diagnosis ; Leptospirosis - genetics ; Livestock ; Melioidosis ; Melioidosis - diagnosis ; Melioidosis - genetics ; Microbiology ; Microorganisms ; Morbidity ; Pneumonia ; Real-Time Polymerase Chain Reaction ; Salmonella ; Strains (organisms) ; Tropical diseases</subject><ispartof>BioMed research international, 2019-01, Vol.2019 (2019), p.1-6</ispartof><rights>Copyright © 2019 Mohammad Ridhuan Mohd Ali et al.</rights><rights>COPYRIGHT 2019 John Wiley & Sons, Inc.</rights><rights>Copyright © 2019 Mohammad Ridhuan Mohd Ali et al. This is an open access article distributed under the Creative Commons Attribution License (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. https://creativecommons.org/licenses/by/4.0</rights><rights>Copyright © 2019 Mohammad Ridhuan Mohd Ali et al. 2019</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c456t-24c1f1b25ca59bf4642525fdbf144bc0b71718045bc276c3a1c28444870866613</cites><orcidid>0000-0002-0034-7624 ; 0000-0002-8626-5848 ; 0000-0002-5888-3526 ; 0000-0002-5867-9816 ; 0000-0001-8999-028X ; 0000-0002-9458-2288 ; 0000-0002-0426-2855</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/PMC6633960/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6633960/$$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/31355287$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Mancianti, Francesca</contributor><creatorcontrib>Ismail, Nabilah</creatorcontrib><creatorcontrib>Yean Yean, Chan</creatorcontrib><creatorcontrib>Harun, Azian</creatorcontrib><creatorcontrib>Aziah, Ismail</creatorcontrib><creatorcontrib>Mustaffa, Khairul Mohd Fadzli</creatorcontrib><creatorcontrib>Ismail, Asmaliza S.</creatorcontrib><creatorcontrib>Goay, Yuan Xin</creatorcontrib><creatorcontrib>Foo, Phiaw Chong</creatorcontrib><creatorcontrib>Lih Huey, Lee</creatorcontrib><creatorcontrib>Mohd Ali, Mohammad Ridhuan</creatorcontrib><creatorcontrib>Phua, Kia Kien</creatorcontrib><title>Duplex TaqMan Hydrolysis Probe-Based Molecular Assay for Simultaneous Detection and Differentiation of Burkholderia pseudomallei and Leptospira spp. DNA</title><title>BioMed research international</title><addtitle>Biomed Res Int</addtitle><description>Melioidosis and leptospirosis, caused by two different bacteria, Burkholderia pseudomallei and Leptospira spp., are potentially fatal infections that share a very similar spectrum of clinical features and cause significant mortality and morbidity in humans and livestock. 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In conclusion, the duplex qPCR assay was sensitive and specific for the detection of B. pseudomallei & Leptospira spp. DNA and is suitable for further analytical and clinical evaluation.</description><subject>Animals</subject><subject>Assaying</subject><subject>Burkholderia pseudomallei</subject><subject>Burkholderia pseudomallei - genetics</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnostic software</subject><subject>Diagnostic systems</subject><subject>DNA</subject><subject>DNA, Bacterial - genetics</subject><subject>Genetic research</subject><subject>Gram-negative bacteria</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Hydrolysis</subject><subject>Infection</subject><subject>Infectious diseases</subject><subject>Leptospira</subject><subject>Leptospira - genetics</subject><subject>Leptospirosis</subject><subject>Leptospirosis - diagnosis</subject><subject>Leptospirosis - genetics</subject><subject>Livestock</subject><subject>Melioidosis</subject><subject>Melioidosis - 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Early detection is important for better clinical consequences. To our knowledge, there is no diagnostic tool available to simultaneously detect and differentiate melioidosis and leptospirosis in humans and animals. In this study, we described a duplex TaqMan probe-based qPCR for the detection of B. pseudomallei and Leptospira spp. DNA. The performance of the assay was evaluated on 20 B. pseudomallei isolates, 23 Leptospira strains, and 39 other microorganisms, as well as two sets of serially diluted reference strains. The duplex qPCR assay was able to detect 0.02 pg (~ 4 copies) Leptospira spp. DNA and 0.2 pg (~ 25.6 copies) B. pseudomallei DNA. No undesired amplification was observed in other microorganisms. In conclusion, the duplex qPCR assay was sensitive and specific for the detection of B. pseudomallei & Leptospira spp. DNA and is suitable for further analytical and clinical evaluation.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Publishing Corporation</pub><pmid>31355287</pmid><doi>10.1155/2019/9451791</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0002-0034-7624</orcidid><orcidid>https://orcid.org/0000-0002-8626-5848</orcidid><orcidid>https://orcid.org/0000-0002-5888-3526</orcidid><orcidid>https://orcid.org/0000-0002-5867-9816</orcidid><orcidid>https://orcid.org/0000-0001-8999-028X</orcidid><orcidid>https://orcid.org/0000-0002-9458-2288</orcidid><orcidid>https://orcid.org/0000-0002-0426-2855</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Assaying Burkholderia pseudomallei Burkholderia pseudomallei - genetics Deoxyribonucleic acid Diagnostic software Diagnostic systems DNA DNA, Bacterial - genetics Genetic research Gram-negative bacteria Health aspects Humans Hydrolysis Infection Infectious diseases Leptospira Leptospira - genetics Leptospirosis Leptospirosis - diagnosis Leptospirosis - genetics Livestock Melioidosis Melioidosis - diagnosis Melioidosis - genetics Microbiology Microorganisms Morbidity Pneumonia Real-Time Polymerase Chain Reaction Salmonella Strains (organisms) Tropical diseases |
| title | Duplex TaqMan Hydrolysis Probe-Based Molecular Assay for Simultaneous Detection and Differentiation of Burkholderia pseudomallei and Leptospira spp. DNA |
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