Publication:
Clinically practical seminested PCR for Burkholderia pseudomallei quantitated by enzyme immunoassay with and without solution hybridization

dc.contributor.authorM. Kunakornen_US
dc.contributor.authorR. B. Markhamen_US
dc.contributor.otherMahidol Universityen_US
dc.date.accessioned2018-07-04T06:59:38Z
dc.date.available2018-07-04T06:59:38Z
dc.date.issued1995-01-01en_US
dc.description.abstractDiagnosis of melioidosis, an infectious disease caused by Burkholderia pseudomallei (formerly Pseudomonas pseudomallei), is made initially by antibody testing, which is not always sensitive or specific. We have developed two seminested PCR protocols combined with enzyme immunoassay (EIA) to detect the conserved ribosomal regulatory region of B. pseudomallei. Both PCRs used one biotinylated primer for capturing PCR products on EIA plates. One system, termed solution hybridization EIA (SHEIA), hybridized PCR products with a digoxigenin-labeled probe in solution. Another system, termed primer-labeled EIA (PLEIA), used a digoxigenin-labeled nested primer to generate products that were directly detected without hybridization. To prevent amplicon contamination, pre-PCR uracil DNA glycosylase treatment or post-PCR UV irradiation was incorporated into each system. By a rapid method of blood sample preparation for PCR, these systems had sensitivities of 75 bacteria per ml for SHEIA and 300 bacteria per ml for PLEIA. No nonspecific amplification of other bacterial DNAs was detected. This seminested PCR coupled with SHEIA or PLEIA fulfills all the requirements for a diagnostic test to be used in developing countries where B. pseudomallei is endemic.en_US
dc.identifier.citationJournal of Clinical Microbiology. Vol.33, No.8 (1995), 2131-2135en_US
dc.identifier.issn00951137en_US
dc.identifier.other2-s2.0-0029020948en_US
dc.identifier.urihttps://repository.li.mahidol.ac.th/handle/20.500.14594/17472
dc.rightsMahidol Universityen_US
dc.rights.holderSCOPUSen_US
dc.source.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=0029020948&origin=inwarden_US
dc.subjectMedicineen_US
dc.titleClinically practical seminested PCR for Burkholderia pseudomallei quantitated by enzyme immunoassay with and without solution hybridizationen_US
dc.typeArticleen_US
dspace.entity.typePublication
mu.datasource.scopushttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=0029020948&origin=inwarden_US

Files

Collections