Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species
Summary For early detection and species differentiation of mycobacteria, polymerase chain reaction (PCR) techniques are currently in wide use. However, individual techniques using amplification of different targets with appropriate primers still have some limitations, which have to be overcome. The...
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| Veröffentlicht in: | Tropical medicine & international health 2003-02, Vol.8 (2), p.150-157 |
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| creator | Bhattacharya, Basudev Karak, Kalpana Ghosal, Alok Gopal Roy, Atanu Das, Shyamal Dandapat, Premanshu Khetawat, Dimple Mondal, Dibya Kanti Bhattacharya, Sujit Chakrabarti, Sekhar |
| description | Summary
For early detection and species differentiation of mycobacteria, polymerase chain reaction (PCR) techniques are currently in wide use. However, individual techniques using amplification of different targets with appropriate primers still have some limitations, which have to be overcome. The ideal technique would use DNA sequences which should be present in all mycobacteria and absent in others and would be able to discriminate one species from the other, as non‐tuberculous mycobacteria (NTM) are on rise in terms of frequency of detection. We developed a multiplex PCR based on amplification of 165, 365 and 541 bp target fragments of unrelated genes, hsp 65 coding for 65 kDa antigen, dnaJ gene of mycobacteria and insertion element IS 6110 of Mycobacterium tuberculosis, respectively. This multiplex PCR was tested over 5 years from 1996 to 2001 with 411 clinical specimens from suspected cases of tuberculosis and mycobacterioses and compared with standard laboratory techniques. The multiplex PCR was positive for 379 cases compared with 280 cases by standard techniques (P |
| doi_str_mv | 10.1046/j.1365-3156.2003.01007.x |
| format | Article |
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For early detection and species differentiation of mycobacteria, polymerase chain reaction (PCR) techniques are currently in wide use. However, individual techniques using amplification of different targets with appropriate primers still have some limitations, which have to be overcome. The ideal technique would use DNA sequences which should be present in all mycobacteria and absent in others and would be able to discriminate one species from the other, as non‐tuberculous mycobacteria (NTM) are on rise in terms of frequency of detection. We developed a multiplex PCR based on amplification of 165, 365 and 541 bp target fragments of unrelated genes, hsp 65 coding for 65 kDa antigen, dnaJ gene of mycobacteria and insertion element IS 6110 of Mycobacterium tuberculosis, respectively. This multiplex PCR was tested over 5 years from 1996 to 2001 with 411 clinical specimens from suspected cases of tuberculosis and mycobacterioses and compared with standard laboratory techniques. The multiplex PCR was positive for 379 cases compared with 280 cases by standard techniques (P < 0.0001). It could distinguish between strains of the M. tuberculosis complex and NTM; the results are comparable with standard techniques. Thus the multiplex PCR can be useful in early detection, species differentiation and epidemiology.</description><identifier>ISSN: 1360-2276</identifier><identifier>EISSN: 1365-3156</identifier><identifier>DOI: 10.1046/j.1365-3156.2003.01007.x</identifier><identifier>PMID: 12581441</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Science Ltd</publisher><subject>Bacterial diseases ; Bacterial Typing Techniques - methods ; Bacterial Typing Techniques - standards ; Biological and medical sciences ; Body Fluids - microbiology ; DNA Primers ; DNA, Bacterial - analysis ; Gene Amplification ; Genes, Bacterial ; Human bacterial diseases ; Humans ; India ; Infectious diseases ; Medical sciences ; multiplex polymerase chain reaction ; mycobacteria ; Mycobacterium - classification ; Mycobacterium - isolation & purification ; Mycobacterium tuberculosis ; Mycobacterium tuberculosis - classification ; Mycobacterium tuberculosis - isolation & purification ; Polymerase Chain Reaction - methods ; Polymerase Chain Reaction - standards ; rapid detection ; Sensitivity and Specificity ; Suppuration - microbiology ; Tropical medicine ; Tuberculosis - diagnosis ; Tuberculosis - microbiology ; Tuberculosis and atypical mycobacterial infections</subject><ispartof>Tropical medicine & international health, 2003-02, Vol.8 (2), p.150-157</ispartof><rights>2003 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4767-6ae10a62aa931cd1ecebb72fe488ee32be9c39ae7f1f08f0c1b5b138b8bd79283</citedby><cites>FETCH-LOGICAL-c4767-6ae10a62aa931cd1ecebb72fe488ee32be9c39ae7f1f08f0c1b5b138b8bd79283</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1046%2Fj.1365-3156.2003.01007.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1046%2Fj.1365-3156.2003.01007.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,777,781,1412,1428,27905,27906,45555,45556,46390,46814</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14549117$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12581441$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bhattacharya, Basudev</creatorcontrib><creatorcontrib>Karak, Kalpana</creatorcontrib><creatorcontrib>Ghosal, Alok Gopal</creatorcontrib><creatorcontrib>Roy, Atanu</creatorcontrib><creatorcontrib>Das, Shyamal</creatorcontrib><creatorcontrib>Dandapat, Premanshu</creatorcontrib><creatorcontrib>Khetawat, Dimple</creatorcontrib><creatorcontrib>Mondal, Dibya Kanti</creatorcontrib><creatorcontrib>Bhattacharya, Sujit</creatorcontrib><creatorcontrib>Chakrabarti, Sekhar</creatorcontrib><title>Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species</title><title>Tropical medicine & international health</title><addtitle>Trop Med Int Health</addtitle><description>Summary
For early detection and species differentiation of mycobacteria, polymerase chain reaction (PCR) techniques are currently in wide use. However, individual techniques using amplification of different targets with appropriate primers still have some limitations, which have to be overcome. The ideal technique would use DNA sequences which should be present in all mycobacteria and absent in others and would be able to discriminate one species from the other, as non‐tuberculous mycobacteria (NTM) are on rise in terms of frequency of detection. We developed a multiplex PCR based on amplification of 165, 365 and 541 bp target fragments of unrelated genes, hsp 65 coding for 65 kDa antigen, dnaJ gene of mycobacteria and insertion element IS 6110 of Mycobacterium tuberculosis, respectively. This multiplex PCR was tested over 5 years from 1996 to 2001 with 411 clinical specimens from suspected cases of tuberculosis and mycobacterioses and compared with standard laboratory techniques. The multiplex PCR was positive for 379 cases compared with 280 cases by standard techniques (P < 0.0001). It could distinguish between strains of the M. tuberculosis complex and NTM; the results are comparable with standard techniques. Thus the multiplex PCR can be useful in early detection, species differentiation and epidemiology.</description><subject>Bacterial diseases</subject><subject>Bacterial Typing Techniques - methods</subject><subject>Bacterial Typing Techniques - standards</subject><subject>Biological and medical sciences</subject><subject>Body Fluids - microbiology</subject><subject>DNA Primers</subject><subject>DNA, Bacterial - analysis</subject><subject>Gene Amplification</subject><subject>Genes, Bacterial</subject><subject>Human bacterial diseases</subject><subject>Humans</subject><subject>India</subject><subject>Infectious diseases</subject><subject>Medical sciences</subject><subject>multiplex polymerase chain reaction</subject><subject>mycobacteria</subject><subject>Mycobacterium - classification</subject><subject>Mycobacterium - isolation & purification</subject><subject>Mycobacterium tuberculosis</subject><subject>Mycobacterium tuberculosis - classification</subject><subject>Mycobacterium tuberculosis - isolation & purification</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polymerase Chain Reaction - standards</subject><subject>rapid detection</subject><subject>Sensitivity and Specificity</subject><subject>Suppuration - microbiology</subject><subject>Tropical medicine</subject><subject>Tuberculosis - diagnosis</subject><subject>Tuberculosis - microbiology</subject><subject>Tuberculosis and atypical mycobacterial infections</subject><issn>1360-2276</issn><issn>1365-3156</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc1u1DAUhSNERUvhFZA3IFgk-NpO7CxYVMNPKxWBUFlbjnMtPHJ-sDPtzNv0UUlmRu2Srnx173fOsXSyjAAtgIrq47oAXpU5h7IqGKW8oECpLLbPsrOHw_P9THPGZHWavUxpTSkVoqxeZKfASgVCwFl2_xlvMQxjh_1EBkcM6fGOJOyTn_wtEtO3BJ3z1i9AtwmTHwNuyTiEXYfRJCT2j_E9iWjs5IeevP-5-vWBuCES384aP2vN_rBYtd45jMv6sJsTH1ak29mhmV0wehNIGnEOTa-yE2dCwtfH9zz7_fXLzeoyv_7x7Wp1cZ1bISuZVwaBmooZU3OwLaDFppHMoVAKkbMGa8trg9KBo8pRC03ZAFeNalpZM8XPs3cH3zEOfzeYJt35ZDEE0-OwSVpyylQN4r8gSGC8ZnwG1QG0cUgpotNj9J2JOw1ULzXqtV7a0ktbeqlR72vU21n65pixaTpsH4XH3mbg7REwyZrgoumtT4-cKEUNIGfu04G78wF3T_6Avvl-tUz8HxNmvK8</recordid><startdate>200302</startdate><enddate>200302</enddate><creator>Bhattacharya, Basudev</creator><creator>Karak, Kalpana</creator><creator>Ghosal, Alok Gopal</creator><creator>Roy, Atanu</creator><creator>Das, Shyamal</creator><creator>Dandapat, Premanshu</creator><creator>Khetawat, Dimple</creator><creator>Mondal, Dibya Kanti</creator><creator>Bhattacharya, Sujit</creator><creator>Chakrabarti, Sekhar</creator><general>Blackwell Science Ltd</general><general>Blackwell Science</general><scope>IQODW</scope><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>7QL</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>200302</creationdate><title>Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species</title><author>Bhattacharya, Basudev ; Karak, Kalpana ; Ghosal, Alok Gopal ; Roy, Atanu ; Das, Shyamal ; Dandapat, Premanshu ; Khetawat, Dimple ; Mondal, Dibya Kanti ; Bhattacharya, Sujit ; Chakrabarti, Sekhar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4767-6ae10a62aa931cd1ecebb72fe488ee32be9c39ae7f1f08f0c1b5b138b8bd79283</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Bacterial diseases</topic><topic>Bacterial Typing Techniques - methods</topic><topic>Bacterial Typing Techniques - standards</topic><topic>Biological and medical sciences</topic><topic>Body Fluids - microbiology</topic><topic>DNA Primers</topic><topic>DNA, Bacterial - analysis</topic><topic>Gene Amplification</topic><topic>Genes, Bacterial</topic><topic>Human bacterial diseases</topic><topic>Humans</topic><topic>India</topic><topic>Infectious diseases</topic><topic>Medical sciences</topic><topic>multiplex polymerase chain reaction</topic><topic>mycobacteria</topic><topic>Mycobacterium - classification</topic><topic>Mycobacterium - isolation & purification</topic><topic>Mycobacterium tuberculosis</topic><topic>Mycobacterium tuberculosis - classification</topic><topic>Mycobacterium tuberculosis - isolation & purification</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Polymerase Chain Reaction - standards</topic><topic>rapid detection</topic><topic>Sensitivity and Specificity</topic><topic>Suppuration - microbiology</topic><topic>Tropical medicine</topic><topic>Tuberculosis - diagnosis</topic><topic>Tuberculosis - microbiology</topic><topic>Tuberculosis and atypical mycobacterial infections</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bhattacharya, Basudev</creatorcontrib><creatorcontrib>Karak, Kalpana</creatorcontrib><creatorcontrib>Ghosal, Alok Gopal</creatorcontrib><creatorcontrib>Roy, Atanu</creatorcontrib><creatorcontrib>Das, Shyamal</creatorcontrib><creatorcontrib>Dandapat, Premanshu</creatorcontrib><creatorcontrib>Khetawat, Dimple</creatorcontrib><creatorcontrib>Mondal, Dibya Kanti</creatorcontrib><creatorcontrib>Bhattacharya, Sujit</creatorcontrib><creatorcontrib>Chakrabarti, Sekhar</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Tropical medicine & international health</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bhattacharya, Basudev</au><au>Karak, Kalpana</au><au>Ghosal, Alok Gopal</au><au>Roy, Atanu</au><au>Das, Shyamal</au><au>Dandapat, Premanshu</au><au>Khetawat, Dimple</au><au>Mondal, Dibya Kanti</au><au>Bhattacharya, Sujit</au><au>Chakrabarti, Sekhar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species</atitle><jtitle>Tropical medicine & international health</jtitle><addtitle>Trop Med Int Health</addtitle><date>2003-02</date><risdate>2003</risdate><volume>8</volume><issue>2</issue><spage>150</spage><epage>157</epage><pages>150-157</pages><issn>1360-2276</issn><eissn>1365-3156</eissn><abstract>Summary
For early detection and species differentiation of mycobacteria, polymerase chain reaction (PCR) techniques are currently in wide use. However, individual techniques using amplification of different targets with appropriate primers still have some limitations, which have to be overcome. The ideal technique would use DNA sequences which should be present in all mycobacteria and absent in others and would be able to discriminate one species from the other, as non‐tuberculous mycobacteria (NTM) are on rise in terms of frequency of detection. We developed a multiplex PCR based on amplification of 165, 365 and 541 bp target fragments of unrelated genes, hsp 65 coding for 65 kDa antigen, dnaJ gene of mycobacteria and insertion element IS 6110 of Mycobacterium tuberculosis, respectively. This multiplex PCR was tested over 5 years from 1996 to 2001 with 411 clinical specimens from suspected cases of tuberculosis and mycobacterioses and compared with standard laboratory techniques. The multiplex PCR was positive for 379 cases compared with 280 cases by standard techniques (P < 0.0001). It could distinguish between strains of the M. tuberculosis complex and NTM; the results are comparable with standard techniques. Thus the multiplex PCR can be useful in early detection, species differentiation and epidemiology.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>12581441</pmid><doi>10.1046/j.1365-3156.2003.01007.x</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Bacterial diseases Bacterial Typing Techniques - methods Bacterial Typing Techniques - standards Biological and medical sciences Body Fluids - microbiology DNA Primers DNA, Bacterial - analysis Gene Amplification Genes, Bacterial Human bacterial diseases Humans India Infectious diseases Medical sciences multiplex polymerase chain reaction mycobacteria Mycobacterium - classification Mycobacterium - isolation & purification Mycobacterium tuberculosis Mycobacterium tuberculosis - classification Mycobacterium tuberculosis - isolation & purification Polymerase Chain Reaction - methods Polymerase Chain Reaction - standards rapid detection Sensitivity and Specificity Suppuration - microbiology Tropical medicine Tuberculosis - diagnosis Tuberculosis - microbiology Tuberculosis and atypical mycobacterial infections |
| title | Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species |
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