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目的制备快速鉴定分枝杆菌菌种的DNA微阵列。方法根据19种分枝杆菌标准株的16SRNA序列设计寡核苷酸探针,制备DNA微阵列,通过反向杂交技术鉴定19种分枝杆菌标准株、9种非分枝杆菌标准株和31株分枝杆菌临床分离株带荧光标记的PCR产物。结果该DNA微阵列与9种非分枝杆菌标准株不杂交,具有分枝杆菌特异性;可将19种分枝杆菌标准株鉴定到群或种,具有较高的分枝杆菌种特异性。应用PCR-SSCP分枝杆菌初步菌种鉴定方法对31株分枝杆菌临床分离株进行初步的鉴定,9株为结核分枝杆菌复合群和22株为非结核分枝杆菌。应用DNA微阵列进一步进行菌种鉴定,9株与探针a杂交阳性,为结核分枝杆菌复合群;2株与探针c杂交阳性,为胞内分枝杆菌;8株与探针d杂交阳性,为堪萨斯分枝杆菌或瘰疬分枝杆菌或猿猴分枝杆菌或胃分枝杆菌;1株与探针k杂交阳性,为戈登分枝杆菌;2株与探针p杂交阳性,为龟分枝杆菌脓肿亚种;2株与探针r杂交阳性,为偶然分枝杆菌;1株与探针s杂交阳性,为草分枝杆菌;2株与探针a和p杂交阳性,为结核分枝杆菌和龟分枝杆菌脓肿亚种复合感染株;1株与探针a和r杂交阳性,为结核分枝杆菌和偶然分枝杆菌复合感染株;3株与该芯片杂交阴性。2株临床分离株经基因测序也证实DNA微阵列鉴定的特异性。结论该DNA微阵列有可能简便、快速、准确地
Objective To prepare DNA microarrays for rapid identification of mycobacterial species. Methods Based on the 16SRNA sequences of 19 mycobacteria strains, oligonucleotide probes were designed and DNA microarrays were prepared. Nineteen mycobacteria strains, nine non-mycobacteria strains and 31 strains were identified by reverse hybridization Mycobacterium clinical isolates with fluorescently labeled PCR product. Results The DNA microarray did not hybridize with 9 non-mycobacterial standard strains and had mycobacterial specificity. Nineteen mycobacteria strains could be identified in groups or species with high mycobacterial species specificity. Thirty-one mycobacteria clinical isolates were initially identified by PCR-SSCP mycobacterial preliminary strain identification method, nine were Mycobacterium tuberculosis complex and 22 were non-tuberculous mycobacteria. DNA microarray was used to further identify the strains. Nine strains were positive for hybridization with probe a, which was Mycobacterium tuberculosis complex. Two strains were positive for probe c and were Mycobacterium intracellulare. Eight strains were hybridized with probe d Positive for Kansas Mycobacterium or Mycobacterium or Mycobacterium or Mycobacterium simonii or Mycobacterium gastris; a positive hybridization with probe k, Mycobacterium Gordon; 2 and probe p hybrid positive for Mycobacterium bovis abscess subspecies; 2 positive hybridization with probe r, which is a chance mycobacterium; 1 and s probe hybridization positive for Mycobacterium phlei; 2 and probe a and p hybridization positive for Mycobacterium tuberculosis and Mycobacterium turtle abscess subspecies complex strains; 1 and probe a and r hybrid positive for the combination of Mycobacterium tuberculosis and incidental mycobacteria; 3 and the chip hybridization negative. Two strains of clinical isolates also confirmed the specificity of DNA microarray by gene sequencing. Conclusion The DNA microarray may be simple, rapid and accurate