本文選題：華支睪吸蟲　切入點：免疫膠體金　出處：《中國熱帶醫(yī)學(xué)》2016年12期

【摘要】：目的建立華支睪吸蟲的免疫膠體金(ICT)和實時熒光定量PCR檢測體系,比較兩者檢測華支睪吸蟲的效果。方法以華支睪吸蟲成蟲抗原包被膠體金,檢測血液樣本中的相應(yīng)抗體,建立華支睪吸蟲ICT檢測方法;以華支睪吸蟲18S r RNA基因作為靶基因,設(shè)計RT-PCR的特異性引物和探針,檢測糞便樣本中的核酸實時熒光定量PCR方法;以兩種檢測體系與商品化的酶聯(lián)免疫(ELISA)檢測試劑盒平行測定比較,考核檢測體系的應(yīng)用價值。結(jié)果對200份華支睪吸蟲重點人群樣本進行檢測,ELISA法檢出的華支睪吸蟲Ig G抗體陽性例數(shù)為20例,陽性率為10.0%;ICT法為16例,陽性率為8.0%,靈敏度為80.0%,特異度為100.0%,符合率為98.0%,ICT和ELISA兩種檢測方法差異無統(tǒng)計學(xué)意義(χ2=2.25,P0.05);RT-PCR法檢出的糞便樣本中華支睪吸蟲核酸陽性例數(shù)為18例,陽性率為9.0%,靈敏度為90.0%,特異度為100.0%,符合率為99.0%,RT-PCR和ELISA兩種檢測方法差異無統(tǒng)計學(xué)意義(χ2=0.05,P0.05)。說明ICT、RT-PCR兩種檢測方法與ELISA的測定結(jié)果比較一致。結(jié)論建立的ICT及RT-PCR兩種檢測方法特異性好、靈敏度高,兩種檢測方法均適用于華支睪吸蟲現(xiàn)場的快速檢測及定量分析。
[Abstract]:Objective to establish an immunoassay system for clonorchiasis sinensis (ICTT) and real-time fluorescent quantitative PCR (PCR) to compare the effect of the two methods in detecting clonorchiasis sinensis. Methods Colloidal gold was coated with adult worm antigen of Clonorchis sinensis to detect the corresponding antibodies in blood samples. A ICT detection method for Clonorchis sinensis was established, and the 18s r RNA gene of Clonorchis sinensis was used as the target gene. The specific primers and probes of RT-PCR were designed to detect the real-time fluorescence quantitative PCR method of nucleic acid in fecal samples. Two detection systems were compared with commercial Elisa Elisa kit. Results among 200 samples of clonorchiasis sinensis detected by Elisa, 20 cases were positive for IgG antibody, and the positive rate was 10.0% ICT method in 16 cases. The positive rate was 8.0%, the sensitivity was 80.0 and the specificity was 100.0.The coincidence rate was 98.0%. There was no significant difference between the two methods (蠂 2 / 2.25P 0.05 / RT-PCR). 18 cases were positive for nucleic acid of Clonorchis sinensis. The positive rate was 9.0%, the sensitivity was 90.0 and the specificity was 100.0.The coincidence rate was 99.0%. There was no significant difference between RT-PCR and ELISA (蠂 ~ 2 / 0.05) (蠂 ~ 2 / 0.05). The results showed that the results of RT-PCR and RT-PCR were consistent with those of ELISA. Conclusion the established ICT and RT-PCR methods have good specificity. The two methods are suitable for rapid detection and quantitative analysis of Clonorchis sinensis.
【作者單位】： 深圳市疾病預(yù)防控制中心;南華大學(xué)公共衛(wèi)生學(xué)院;
【基金】：國家衛(wèi)生行業(yè)科研專項(No.201502021)
【分類號】：R532.23;R440

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