Fries J W, Patel R J, Piessens W F, Wirth D F
Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115.
Mol Cell Probes. 1990 Apr;4(2):87-105. doi: 10.1016/0890-8508(90)90011-n.
Differential diagnosis of Mycobacterium tuberculosis, M. avium, and other mycobacteria remains a lengthy process. Recently, the use of DNA probes has been proposed as a new approach for a more specific and rapid diagnosis. Here, we report the cloning and sequencing of a genus-specific probe for Mycobacterium and a species-specific M. avium probe. The genus-specific probe hybridizes with DNA from nine ATCC type strains and 13 isolates of mycobacteria but not to non-mycobacterial DNA. In addition, the cloned fragment could also be amplified by polymerase chain reaction (PCR) in DNa of ten different mycobacterial type strains. The M. avium specific probe hybridizes strongly to sequences amplified in M. avium but not other mycobacterial or non-mycobacterial DNA. Amplification of the target sequence by PCR allowed the detection of 1 fg of all mycobacterial DNA tested for the genus-specific probe and 1 fg of M. avium DNA for the species-specific probe.
结核分枝杆菌、鸟分枝杆菌及其他分枝杆菌的鉴别诊断仍是一个漫长的过程。最近,有人提出使用DNA探针作为一种更特异、快速诊断的新方法。在此,我们报告了一种分枝杆菌属特异性探针和一种鸟分枝杆菌种特异性探针的克隆及测序。该属特异性探针能与9株美国典型培养物保藏中心(ATCC)标准菌株及13株分枝杆菌分离株的DNA杂交,但不与非分枝杆菌DNA杂交。此外,克隆片段还可通过聚合酶链反应(PCR)在10种不同分枝杆菌标准菌株的DNA中进行扩增。鸟分枝杆菌特异性探针能与鸟分枝杆菌中扩增的序列强烈杂交,但不与其他分枝杆菌或非分枝杆菌DNA杂交。通过PCR扩增靶序列,对于属特异性探针,可检测到1 fg的所有受试分枝杆菌DNA;对于种特异性探针,可检测到1 fg的鸟分枝杆菌DNA。