Lage A P, Fauconnier A, Burette A, Glupczynski Y, Bollen A, Godfroid E
Service de Genetique Appliquee, Universite Libre de Bruxelles, Belgium.
J Clin Microbiol. 1996 Mar;34(3):530-3. doi: 10.1128/jcm.34.3.530-533.1996.
A very simple, practical, sensitive, and specific colorimetric hybridization assay for detecting amplified Helicobacter pylori DNA is described. This assay, which combines a sensitive sandwich DNA hybridization reaction and a colorimetric protocol similar to those used in conventional enzyme immunoassays, was shown to be suitable for detecting H. pylori-infected gastric biopsy specimens and for monitoring the eradication of the pathogen after treatment. The specificity and sensitivity of the colorimetric hybridization assay were tested by assaying 27 H. pylori strains (4 reference and 23 clinical isolates), 9 strains of other Helicobacter spp. or Campylobacter spp., and 11 clinical isolates of other urease-positive bacteria. The likelihood of H. pylori detection in gastric biopsy specimens by the colorimetric hybridization assay was evaluated with 23 H. pylori-positive and 41 H. pylori-negative biopsy specimens on the basis of positive and negative results, respectively, of culture, rapid urease test, histological examination, and PCR. Biopsy specimens from 33 treated patients, endoscopied 4 to 8 weeks after the end of treatment, were also tested. All H. pylori strains showed positive results in the colorimetric hybridization assay, presenting optical densities at 450 nm (OD450S) of > or = 3.0. None of the other Helicobacter spp., Campylobacter spp., or the clinical isolates of other urease-positive bacteria showed OD450S equal to or greater than the cutoff (mean OD450 cutoff, 0.208). The colorimetric hybridization assay detected all 23 H. pylori-positive biopsy specimens (mean OD450, 2.910 +/- 0.295), while none of the H. pylori-negative biopsy specimens was shown to be positive in the assay (mean OD450, 0.108 +/- 0.025). H. pylori was considered to be not eradicated from three of the posttreatment biopsy specimens by culture, rapid urease test, histological examination, and PCR. They were all positive by the colorimetric hybridization assay, and their OD450S were > or = 3.0. The colorimetric hybridization assay also detected two other H. pylori-positive patients. Specimens from these two patients had negative culture, rapid urease test, and histology results, and a specimen from one of them also tested negative by PCR. These results indicate that the colorimetric hybridization assay is a suitable method both for the diagnosis of H. pylori in biopsy specimens and for the follow-up of patients after the end of treatment.
本文描述了一种用于检测扩增的幽门螺杆菌DNA的非常简单、实用、灵敏且特异的比色杂交检测方法。该检测方法结合了灵敏的夹心DNA杂交反应和类似于传统酶免疫分析中使用的比色方案，被证明适用于检测幽门螺杆菌感染的胃活检标本以及监测治疗后病原体的根除情况。通过检测27株幽门螺杆菌菌株（4株参考菌株和23株临床分离株）、9株其他幽门螺杆菌属或弯曲菌属菌株以及11株其他脲酶阳性细菌的临床分离株，对比色杂交检测方法的特异性和灵敏性进行了测试。基于培养、快速脲酶试验、组织学检查和PCR的阳性和阴性结果，分别用23份幽门螺杆菌阳性和41份幽门螺杆菌阴性的活检标本评估了比色杂交检测方法在胃活检标本中检测幽门螺杆菌的可能性。还对33例治疗后患者的活检标本进行了检测，这些患者在治疗结束后4至8周接受了内镜检查。所有幽门螺杆菌菌株在比色杂交检测中均显示阳性结果，在450nm处的光密度（OD450S）≥3.0。其他幽门螺杆菌属、弯曲菌属菌株或其他脲酶阳性细菌的临床分离株均未显示OD450S等于或大于临界值（平均OD450临界值为0.208）。比色杂交检测方法检测出了所有23份幽门螺杆菌阳性的活检标本（平均OD450为2.910±0.295），而所有幽门螺杆菌阴性的活检标本在该检测中均未显示为阳性（平均OD450为0.108±0.025）。通过培养、快速脲酶试验、组织学检查和PCR，认为3份治疗后活检标本中的幽门螺杆菌未被根除。它们在比色杂交检测中均为阳性，且OD450S≥3.0。比色杂交检测方法还检测出另外两名幽门螺杆菌阳性患者。这两名患者的标本培养、快速脲酶试验和组织学结果均为阴性，其中一名患者的标本PCR检测也为阴性。这些结果表明，比色杂交检测方法是一种适用于诊断活检标本中幽门螺杆菌以及治疗结束后患者随访的方法。
