Sow A, Sidibé I, Desquesnes M, Bengaly Z, Pangui L J
Laboratoire National d'Elevage, 03 BP. 7026 Ouagadougou 03, Burkina Faso.
Trop Biomed. 2006 Jun;23(1):123-9.
PCR-ELISA was set up to detect strains of Trypanosoma congolense type savannah (TCS) in field samples of buffy coats. Results of PCR-ELISA and PCR were compared and the effectiveness of both techniques was also compared with the Murray's method for the detection of TCS in 257 bovine buffy coats. The PCR products were labelled with digoxigenin (DIG-dUTP) during amplification cycles of the repetitive satellite DNA. A biotinylated DNA capture probe was used to detect the PCR products by ELISA in streptavidin coated microplates. Both the PCR-ELISA and PCR were more sensitive and more specific than the Murray's method. Of the 257 buffy coats analysed by the three techniques, PCR-ELISA and PCR detected TCS in 98 and 97 buffy coats respectively, whereas the Murray's method detected only 39 samples. PCR-ELISA and PCR had almost the same sensitivity and specificity. PCR-ELISA and PCR respectively detected TCS in 39.2% and 38.6% in all the 334 samples analysed by both techniques in this study.
建立了PCR-ELISA法来检测血沉棕黄层野外样本中的刚果锥虫草原亚种（TCS）菌株。比较了PCR-ELISA法和PCR法的结果，还将这两种技术的有效性与Murray氏法检测257份牛血沉棕黄层中TCS的有效性进行了比较。在重复性卫星DNA的扩增循环过程中，用洋地黄毒苷（地高辛标记的三磷酸尿苷）标记PCR产物。使用生物素化的DNA捕获探针，通过酶联免疫吸附测定法在链霉抗生物素蛋白包被的微孔板中检测PCR产物。PCR-ELISA法和PCR法都比Murray氏法更灵敏、更特异。用这三种技术分析的257份血沉棕黄层样本中，PCR-ELISA法和PCR法分别在98份和97份血沉棕黄层样本中检测到了TCS，而Murray氏法仅检测到39份样本。PCR-ELISA法和PCR法的灵敏度和特异性几乎相同。在本研究中用这两种技术分析的所有334份样本中，PCR-ELISA法和PCR法分别在39.2%和38.6%的样本中检测到了TCS。
