Eisenach K D, Crawford J T, Bates J H
Medical Research Service, John L. McClellan Memorial Veterans Hospital, Little Rock, Arkansas.
J Clin Microbiol. 1988 Nov;26(11):2240-5. doi: 10.1128/jcm.26.11.2240-2245.1988.
Three cloned segments of Mycobacterium tuberculosis DNA which are promising as clinical probes were identified. An MboI digest of DNA from a clinical isolate of M. tuberculosis was cloned into bacteriophage M13. To identify recombinants specific for the M. tuberculosis complex, plaque lifts were hybridized with M. bovis and M. kansasii DNA. Recombinants which selectively hybridized with M. bovis DNA were characterized by probing slot blots and restriction digests of DNA from various mycobacteria. Three recombinants that did not hybridize to a significant extent with DNA from nontuberculous mycobacteria were identified. These three probes are of special interest because they are each repeated multiple (10 to 16) times in the M. tuberculosis chromosome. These probes were also shown to be useful for fingerprinting strains for epidemiological studies.
鉴定出了三个有望用作临床探针的结核分枝杆菌DNA克隆片段。将来自结核分枝杆菌临床分离株的DNA经MboI酶切后克隆到噬菌体M13中。为了鉴定结核分枝杆菌复合群特异性的重组体，将噬菌斑转移到硝酸纤维素膜上，与牛分枝杆菌和堪萨斯分枝杆菌DNA进行杂交。通过对来自各种分枝杆菌的DNA进行狭缝印迹杂交和限制性酶切分析，对与牛分枝杆菌DNA选择性杂交的重组体进行了鉴定。鉴定出三个与非结核分枝杆菌DNA没有明显杂交的重组体。这三个探针特别令人感兴趣，因为它们在结核分枝杆菌染色体中各自重复多次（10至16次）。这些探针还被证明可用于对菌株进行指纹图谱分析以开展流行病学研究。
