Gonzales J L, Loza A, Chacon E
Laboratorio de Investigación y Diagnóstico Veterinario "LIDIVET", Av. Ejercito Nacional 153, P.O. Box 29, Santa Cruz, Bolivia.
Vet Parasitol. 2006 Mar 15;136(2):119-26. doi: 10.1016/j.vetpar.2005.10.024. Epub 2005 Dec 15.
There are several T. vivax specific primers developed for PCR diagnosis. Most of these primers were validated under different DNA extraction methods and study designs leading to heterogeneity of results. The objective of the present study was to validate PCR as a diagnostic test for T. vivax trypanosomosis by means of determining the test sensitivity of different published specific primers with different sample preparations. Four different DNA extraction methods were used to test the sensitivity of PCR with four different primer sets. DNA was extracted directly from whole blood samples, blood dried on filter papers or blood dried on FTA cards. The results showed that the sensitivity of PCR with each primer set was highly dependant of the sample preparation and DNA extraction method. The highest sensitivities for all the primers tested were determined using DNA extracted from whole blood samples, while the lowest sensitivities were obtained when DNA was extracted from filter paper preparations. To conclude, the obtained results are discussed and a protocol for diagnosis and surveillance for T. vivax trypanosomosis is recommended.
已经开发出几种用于PCR诊断的间日疟原虫特异性引物。这些引物大多在不同的DNA提取方法和研究设计下进行了验证,导致结果存在异质性。本研究的目的是通过确定不同已发表的特异性引物在不同样品制备情况下的检测灵敏度,来验证PCR作为间日疟原虫锥虫病诊断检测方法的有效性。使用四种不同的DNA提取方法,对四组不同的引物进行PCR灵敏度测试。DNA直接从全血样本、滤纸上干燥的血液或FTA卡上干燥的血液中提取。结果表明,每组引物的PCR灵敏度高度依赖于样品制备和DNA提取方法。所有测试引物的最高灵敏度是使用从全血样本中提取的DNA测定的,而从滤纸制备物中提取DNA时灵敏度最低。总之,对获得的结果进行了讨论,并推荐了间日疟原虫锥虫病的诊断和监测方案。