DNA DIVERSITY AMONG CLINICAL ISOLATES OF HELICOBACTER-PYLORI DETECTED BY PCR-BASED RAPD FINGERPRINTING
DNA DIVERSITY AMONG CLINICAL ISOLATES OF HELICOBACTER-PYLORI DETECTED BY PCR-BASED RAPD FINGERPRINTING
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DOI：
10.1093/nar/20.19.5137
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发表时间：
1992-10-11
影响因子：
14.9
通讯作者：
BERG, DE
中科院分区：
文献类型：
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作者：
AKOPYANZ, N;BUKANOV, NO;BERG, DE
应用RAPD（或AP-PCR） DNA指纹图谱方法对幽门螺杆菌临床分离株进行区分。幽门螺杆菌是一种长期携带与胃炎、消化性溃疡和胃癌相关的细菌。这种方法使用任意选择的寡核苷酸，从它们偶然匹配或几乎匹配的基因组位点启动DNA合成。大多数含有大于或等于60% G+C的10-nt引物产生了多达15个突出片段的菌株特异性阵列，大多数较长的（大于或等于17-nt）引物也是如此，而大多数含有50% G+C的10-nt引物则没有。64株独立的幽门螺杆菌分离株（其中60株来自同一医院的患者）均可通过单一RAPD引物进行区分，这表明该物种具有高度的DNA序列多样性。相比之下，从最初的和后续的活检中分离出的分离株在三个检测的病例中都是难以区分的。
The RAPD (or AP-PCR) DNA fingerprinting method was used to distinguish among clinical isolates of Helicobacter pylori, a bacterium whose long term carriage is associated with gastritis, peptic ulcers and gastric carcinomas. This method uses arbitrarily chosen oligonucleotides to prime DNA synthesis from genomic sites to which they are fortuitously matched, or almost matched. Most 10-nt primers with greater-than-or-equal-to 60% G+C yielded strain-specific arrays of up to 15 prominent fragments, as did most longer (greater-than-or-equal-to 17-nt) primers, whereas most 10-nt primers with 50% G+C did not. Each of 64 independent H.pylori isolates, 60 of which were from patients in the same hospital, was distinguishable with a single RAPD primer, which suggests a high level of DNA sequence diversity within this species. In contrast, isolates from initial and followup biopsies were indistinguishable in each of three cases tested.