Cited 36 times in

Development of a One-Step Multiplex PCR Assay for Differential Detection of Major Mycobacterium Species

DC Field Value Language
dc.contributor.author신성재-
dc.contributor.author용동은-
dc.date.accessioned2018-07-20T08:03:12Z-
dc.date.available2018-07-20T08:03:12Z-
dc.date.issued2017-
dc.identifier.issn0095-1137-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/160789-
dc.description.abstractThe prevalence of tuberculosis continues to be high, and nontuberculous mycobacterial (NTM) infection has also emerged worldwide. Moreover, differential and accurate identification of mycobacteria to the species or subspecies level is an unmet clinical need. Here, we developed a one-step multiplex PCR assay using whole-genome analysis and bioinformatics to identify novel molecular targets. The aims of this assay were to (i) discriminate between the Mycobacterium tuberculosis complex (MTBC) and NTM using rv0577 or RD750, (ii) differentiate M. tuberculosis (M. tuberculosis) from MTBC using RD9, (iii) selectively identify the widespread M. tuberculosis Beijing genotype by targeting mtbk_20680, and (iv) simultaneously detect five clinically important NTM (M. avium, M. intracellulare, M. abscessus, M. massiliense, and M. kansasii) by targeting IS1311, DT1, mass_3210, and mkan_rs12360 An initial evaluation of the multiplex PCR assay using reference strains demonstrated 100% specificity for the targeted Mycobacterium species. Analytical sensitivity ranged from 1 to 10 pg for extracted DNA and was 103 and 104 CFU for pure cultures and nonhomogenized artificial sputum cultures, respectively, of the targeted species. The accuracy of the multiplex PCR assay was further evaluated using 55 reference strains and 94 mycobacterial clinical isolates. Spoligotyping, multilocus sequence analysis, and a commercial real-time PCR assay were employed as standard assays to evaluate the multiplex PCR assay with clinical M. tuberculosis and NTM isolates. The PCR assay displayed 100% identification agreement with the standard assays. Our multiplex PCR assay is a simple, convenient, and reliable technique for differential identification of MTBC, M. tuberculosis, M. tuberculosis Beijing genotype, and major NTM species.-
dc.description.statementOfResponsibilityopen-
dc.languageEnglish-
dc.publisherAmerican Society for Microbiology-
dc.relation.isPartOfJOURNAL OF CLINICAL MICROBIOLOGY-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.rightshttps://creativecommons.org/licenses/by-nc-nd/2.0/kr/-
dc.subject.MESHHumans-
dc.subject.MESHMultilocus Sequence Typing/methods-
dc.subject.MESHMultiplex Polymerase Chain Reaction/methods*-
dc.subject.MESHMycobacterium Infections, Nontuberculous/diagnosis*-
dc.subject.MESHMycobacterium Infections, Nontuberculous/microbiology-
dc.subject.MESHMycobacterium tuberculosis/genetics-
dc.subject.MESHMycobacterium tuberculosis/isolation & purification*-
dc.subject.MESHNontuberculous Mycobacteria/genetics-
dc.subject.MESHNontuberculous Mycobacteria/isolation & purification*-
dc.subject.MESHRNA, Ribosomal, 16S/genetics-
dc.subject.MESHTuberculosis, Pulmonary/diagnosis*-
dc.subject.MESHTuberculosis, Pulmonary/microbiology-
dc.titleDevelopment of a One-Step Multiplex PCR Assay for Differential Detection of Major Mycobacterium Species-
dc.typeArticle-
dc.contributor.collegeCollege of Medicine-
dc.contributor.departmentDept. of Microbiology-
dc.contributor.googleauthorHansong Chae-
dc.contributor.googleauthorSeung Jung Han-
dc.contributor.googleauthorSu-Young Kim-
dc.contributor.googleauthorChang-Seok Ki-
dc.contributor.googleauthorHee Jae Huh-
dc.contributor.googleauthorDongeun Yong-
dc.contributor.googleauthorWon-Jung Koh-
dc.contributor.googleauthorSung Jae Shin-
dc.identifier.doi10.1128/JCM.00549-17-
dc.contributor.localIdA02114-
dc.contributor.localIdA02423-
dc.relation.journalcodeJ01325-
dc.identifier.eissn1098-660X-
dc.identifier.pmid28659320-
dc.subject.keywordBeijing genotype-
dc.subject.keywordMycobacterium abscessus complex-
dc.subject.keywordMycobacterium avium complex-
dc.subject.keywordMycobacterium kansasii-
dc.subject.keywordMycobacterium tuberculosis-
dc.subject.keywordmultiplex PCR-
dc.subject.keywordnontuberculous mycobacteria-
dc.contributor.alternativeNameShin, Sung Jae-
dc.contributor.alternativeNameYong, Dong Eun-
dc.contributor.affiliatedAuthorShin, Sung Jae-
dc.contributor.affiliatedAuthorYong, Dong Eun-
dc.citation.volume55-
dc.citation.number9-
dc.citation.startPage2736-
dc.citation.endPage2751-
dc.identifier.bibliographicCitationJOURNAL OF CLINICAL MICROBIOLOGY, Vol.55(9) : 2736-2751, 2017-
dc.identifier.rimsid60675-
dc.type.rimsART-
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers

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