Evaluation of culture and PCR-based assay for detection of Mycobacterium tuberculosis from sputum collected and stored on filter paper.

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dc.contributor.authorTansuphasiri, Uen_US
dc.contributor.authorChinrat, Ben_US
dc.contributor.authorRienthong, Sen_US
dc.date.accessioned2009-05-27T16:48:30Z
dc.date.available2009-05-27T16:48:30Z
dc.date.issued2001-12-04en_US
dc.descriptionThe Southeast Asian Journal of Tropical Medicine and Public Health.en_US
dc.description.abstractWe evaluated the use of culture and PCR-based assay for the direct detection of Mycobacterium tuberculosis (MTB) from sputum collected and stored on filter paper at room temperature for 5 days; the results were compared with those of staining and conventional culture of fresh sputum before storage (the 'gold standard'). Out of 231 sputum specimens examined, MTB was recovered from 124 samples by culture before storage. The culture positivity rate was significantly decreased to 70% after 5 days storage. For PCR assay, a fragment of 377 bp of the IS6110 sequence was amplified and detected using three methods: first PCR product combined with agarose gel electrophoresis (AGE); first PCR product with dot blot hybridization (DBH); nested PCR with AGE. Compared with culture, the sensitivity, specificity, and efficiency for first PCR with AGE were 71.8, 100 and 84.9% respectively; PCR with DBH gave results of 89.5, 96.3 and 92.6% respectively; the same values for nested PCR were 96.0, 97.2, and 96.5% respectively. Of these three methods, nested PCR gave excellent sensitivity and specificity with no significant difference (p = 0.727) from conventional culture. The storage of sputum on filter paper and storage at room temperature for 5 days had no apparent effect on the performance of nested PCR. We propose that this collection and storage method be considered for transporting sputum specimens from peripheral health centers or from the field; specimens may be sent by post to a central point for both culture and PCR analysis by trained technicians supervised in accordance with a well-established quality control system.en_US
dc.description.affiliationDepartment of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, Thailand. unchalee@loxley.co.then_US
dc.identifier.citationTansuphasiri U, Chinrat B, Rienthong S. Evaluation of culture and PCR-based assay for detection of Mycobacterium tuberculosis from sputum collected and stored on filter paper. The Southeast Asian Journal of Tropical Medicine and Public Health. 2001 Dec; 32(4): 844-55en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/36056
dc.language.isoengen_US
dc.source.urihttps://www.tm.mahidol.ac.th/seameo/2001-32-4/26-844-855.pdfen_US
dc.subject.meshCulture Techniquesen_US
dc.subject.meshHumansen_US
dc.subject.meshMycobacterium tuberculosis --isolation & purificationen_US
dc.subject.meshPaperen_US
dc.subject.meshPolymerase Chain Reaction --methodsen_US
dc.subject.meshSpecimen Handlingen_US
dc.subject.meshSputum --microbiologyen_US
dc.subject.meshThailanden_US
dc.subject.meshTuberculosis --microbiologyen_US
dc.titleEvaluation of culture and PCR-based assay for detection of Mycobacterium tuberculosis from sputum collected and stored on filter paper.en_US
dc.typeEvaluation Studiesen_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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