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Differentiation of mycobacteria in sputa by duplex polymerase chain reaction for mycobacterial hsp65 gene

Cited 8 time in Web of Science Cited 9 time in Scopus
Authors
Kim, Bum-Joon; Park, Joo-Hee; Lee, Seoung-Ae; Kim, Hong; Cha, Chang-Yong; Kook, Yoon-Hoh; Kim, Eui-Chong; Joo, Sei Ick; Lee, Jae Seok; Yim, Jae-Joon
Issue Date
2008-07-26
Publisher
Elsevier
Citation
Diagn Microbiol Infect Dis. 2008 Oct;62(2):193-8. Epub 2008 Jul 23.
Keywords
AdultAgedAged, 80 and overBacterial Proteins/*genetics/metabolismChaperonin 60Chaperonins/*genetics/metabolismDNA, Bacterial/analysis/isolation & purificationDiagnosis, DifferentialFemaleHumansMaleMiddle AgedMycobacterium/*classification/genetics/isolation & purificationMycobacterium tuberculosis/classification/genetics/isolation & purificationPolymerase Chain Reaction/*methodsSensitivity and SpecificitySequence Analysis, DNASpecies SpecificitySputum/*microbiologyTuberculosis, Pulmonary/*diagnosis/microbiology
Abstract
Early differentiation of mycobacteria in sputa is crucial. This study was set to evaluate the usefulness of a newly developed duplex polymerase chain reaction (PCR) for hsp65 gene-based method in differentiating mycobacteria in sputum with a positive acid-fast bacilli (AFB) smear before culturing. One hundred forty-seven sputa with positive AFB smear were included for the analysis. Mycobacterial species were identified using a newly developed duplex PCR for hsp65 gene followed by a nested PCR-direct sequencing and the conventional colony-based method. Final decision of mycobacterial species were made based on 1) results of species identification based on mycobacterial colonies or 2) results of species identification of other sputa from the same patients and clinical findings. The duplex PCR-based method correctly identified 83.2% sputa from tuberculosis patients and 82.2% sputa from nontuberculous mycobacteria patients, whereas the colony-based method correctly identified 86.1% and 77.8%, respectively. Sensitivity and specificity of the colony-based method for Mycobacterium tuberculosis were 86.1% and 100%, respectively, whereas those of the duplex PCR-based method were 83.2% and 95.6%, respectively. The duplex PCR-based method, to differentiate mycobacterial species in sputa, produced comparable results as those of the colony-based identification method.
ISSN
0732-8893 (Print)
0732-8893 (Linking)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18653300

http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T60-4T24FPV-3-5&_cdi=5016&_user=168665&_orig=search&_coverDate=10%2F31%2F2008&_sk=999379997&view=c&wchp=dGLbVlz-zSkWb&md5=2eb7b300b2c070fa64b0e0901e94e6b5&ie=/sdarticle.pdf

http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T60-4T24FPV-3-5&_cdi=5016&_user=168665&_orig=search&_coverDate=10%2F31%2F2008&_sk=999379997&view=c&wchp=dGLzVlz-zSkzV&md5=2eb7b300b2c070fa64b0e0901e94e6b5&ie=/sdarticle.pdf

https://hdl.handle.net/10371/63089
DOI
https://doi.org/10.1016/j.diagmicrobio.2008.05.013
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College of Medicine/School of Medicine (의과대학/대학원)Laboratory Medicine (검사의학전공)Journal Papers (저널논문_검사의학전공)
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