Real-time PCR检测麻风患者治疗前后及少菌型石蜡标本中麻风菌DNA的价值

发布时间：2018-08-25 16:18

【摘要】：目的应用已建立的实时定量荧光PCR(real-time PCR)方法检测少菌型麻风(PB)患者的石蜡标本,探讨该方法对BI为0患者的辅助诊断价值。方法从石蜡标本中提取的DNA进行麻风菌重复序列的特异性real-time PCR检测,包括短程联合化疗(MDT)治疗前后标本共92例;2008年和2013年收集的界限类偏瘤型麻风(BL)石蜡标本分别为7例和8例。并检测70例临床诊断PB和未定类麻风,同时将RT-PCR检测结果与病理分析结果进行比较。结果 92例(LL:19例;BL:20例;BB:20例;BT:16例;TT:17例)不同型别麻风患者的石蜡标本检测阳性率治疗前、治疗后分别为100.00%,100.00%,90.00%,87.50%,50.00%与40.00%,40.00%,0,0,22.22%。比较保存5年前后15例BL患者石蜡标本,检测结果差异无统计学意义。70例临床诊断PB和未定类麻风病患者的石蜡标本real-time PCR检测阳性率为71.43%(50/70),病理检测与临床符合率为51.43%(36/70),两者的阳性检测率差异有统计学意义(P0.05)。结论建立Taq Man(水解探针)技术的real-time PCR方法可以快速、特异、灵敏的检测石蜡标本中的麻风菌DNA,提高抗酸染色阴性标本的检测阳性率,对临床诊断的指导意义重大。
[Abstract]:Objective to detect the paraffin samples of leprosy patients with leprosy by using the established real-time quantitative PCR (real-time PCR) method and to explore the value of this method in the auxiliary diagnosis of BI 0 patients. Methods DNA extracted from paraffin wax samples was detected by specific real-time PCR of repeated sequences of leprosy bacteria. 92 cases were collected before and after (MDT) treatment, and 7 cases and 8 cases of leprosy (BL) were collected in 2008 and 2013, respectively. 70 cases of PB and undetermined leprosy were detected, and the results of RT-PCR were compared with pathological analysis. Results before treatment, the positive rates of paraffin wax specimens were 100.000.100.0090.90% and 87.50%, 50.00% and 40.0040.022.2220%, respectively, in 92 cases of leprosy patients with different types of leprosy (LL:19: 20 cases, BBW 20 cases, BT: 16 cases, TTW 17 cases). The positive rates of paraffin wax specimens from different types of leprosy patients were 100.000.100.000.90.0050.0050.00% and 40.0040.00022.2220, respectively. The paraffin specimens of 15 patients with BL were compared before and after 5 years of preservation. The positive rate of real-time PCR was 71.43% (50 / 70) in paraffin wax specimens and 51.43% (36 / 70) in clinicopathological diagnosis of PB and leprosy like leprosy. The difference between them was statistically significant (P0.05). Conclusion the real-time PCR method of Taq Man (hydrolysis probe can be used to detect leprosy DNA, in paraffin wax samples quickly, specifically and sensitively. It is of great significance for clinical diagnosis to improve the positive rate of acid-fast staining negative samples.
【作者单位】：首都医科大学附属北京友谊医院北京热带医学研究所;云南省文山州皮肤病防治所;
【基金】：首都医科大学热带病防治研究重点实验室开放研究课题资助
【分类号】：R755

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