Identification of Chlamydial species in crocodiles and chickens by PCR-HRM curve analysis.
作者： T. Robertson , S. Bibby , D. O’Rourke , T. Belfiore , R. Agnew-Crumpton
DOI: 10.1016/J.VETMIC.2010.04.007
关键词: Genetics 、 Sequence analysis 、 Chlamydiales 、 Polymerase chain reaction 、 Amplicon 、 16S ribosomal RNA 、 High Resolution Melt 、 Biology 、 Chlamydophila 、 Chlamydiaceae
摘要: Recently, a PCR protocol (16SG), targeting 16S rRNA gene coupled with high resolution melt (HRM) curve analysis was developed in our laboratory and shown to reliably detect identify the seven different Chlamydiaceae spp. In this study, potential of method assessed for detection differentiation Chlamydiosis clinical specimens. Of total number 733 specimens from range animal species, 219 (30%) were found positive by 16SG PCR. When sufficient amount DNA available (64 submissions), amplicons generated subjected HRM results compared that nucleotide sequencing. all instances, infecting genotyped according identity its sequence reference species. Analysis curves sequences also revealed occurrence Chlamydophila-like, Parachlamydia-like variant Chlamydophila psittaci chickens. These reveal PCR-HRM rapid simultaneous identification animals demonstrate capacity system new during routine diagnostic testings.
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