摘要
[目的]探讨PCR-膜片RDB(recersedotblot)技术检测石蜡包埋组织中的结核杆菌的方法。[方法]合成检测结核杆菌的寡核苷酸探针并制作膜芯片,扩增结核分支杆菌插入序列IS6110基因片段,利用PCR-膜片RDB技术,检测12株结核分支杆菌临床分离株、2株大肠杆菌,120例结核病人及38例非结核病人石蜡包埋组织中结核杆菌。石蜡组织的膜片检测结果与抗酸染色法和PCR检测结果比较。[结果]12株扩增阳性的结核分支杆菌,膜片检测11株杂交阳性,2株大肠杆菌和阴性对照结果均为阴性。120例结核病人石蜡组织标本,抗酸染色,PCR和PCR-膜片RDB技术检测灵敏度分别为43.3%(52/120),76.7%(92/120)和87.5%(105/120),三者比较差异有统计学意义(P﹤0.01)。38例非结核病人石蜡组织标本,抗酸染色,PCR和PCR-膜片RDB技术检测特异度分别为100%(38/38),94.7%(36/38)和100%(38/38),三者比较差异无统计学意义(P﹥0.05)。[结论]PCR-膜片RDB技术有助于提高石蜡包埋组织中结核杆菌检出率。
[Objective] To study detection of Mycobacterium tuberculosis in paraffin-embedded tissues specimens by PCR-reverse dot blot membrane chip. [Methods ] Oligonucleotide probe of detecting Mycobaeterium tuberculosis were prepared and gene membrane chips were built. IS6110 gene of Mycobacterium tuberculosis was anlplified. Mycobaeterium tubereulosis in 12 M. tuberculosis clinical isolates, 2 E.coli strains, paraffin-embedded tissues specimens of 120 patients with tuberculosis and 38 cases of nontuberculosis were detected by PCR-reverse dot blot membrane chip. The results of paraffin-embedded tissues specimens were compared to the results of the smear fast-acid staining and PCR. [ Results] 12 M. tuberculosis clinical isolates were positive by PCR, among then 11 were positive by PCR-reverse dot blot membrane chip. 2 E.coli strains and contro] were negative by PCR-reverse dot blot membrane chip. 120 paraffin-embedded tissues specimens of patients with tuberculoSiS were detected by the smear fast-acid staining, PCR and PCR-reverse dot blot membrane chip, the sensitivity was 43.3% （52/120）, 76.7% （92/120） and 87.5% （105/120）, respectively. There was significant difference between the sensitivity （P 〈 0.01）, 38 paraffin-embedded tissues specimens of nontuberculosis were detected by the smear fast-acid staining, PCR and PCR-reverse dot blot membrane chip, the specificity was 100% （38/38）, 94.7% （36/38） and 100% （38/38）, respectively. There Was no significant difference between the sensitivity （P 〉 0.05）. [Conclusion] The technolog of PCR-reverse dot blot membrane chip helps to increase the detection rate of Mycobacterium tuberculosis in paraffin-embedded tissues specimens.
出处
《现代预防医学》
CAS
北大核心
2008年第4期766-768,共3页
Modern Preventive Medicine
基金
广西科技厅资助项目(桂科攻0235024-27,0330004-11)
广西高校百名中青年学科带头人资助计划
关键词
反向杂交膜片
石蜡包埋组织
结核杆菌
Reverse dot blot membrane
Paraffin-embedded tissues, Mycobacterium tuberculosis
