Comparison of polymerase chain reaction methods for the detection of Theileria equi infection using whole blood compared with pre-extracted DNA samples as PCR templates
Comparison of polymerase chain reaction methods for the detection of Theileria equi infection using whole blood compared with pre-extracted DNA samples as PCR templates
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DOI：
10.1007/s11250-007-9025-1
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发表时间：
2007-06-01
影响因子：
1.7
通讯作者：
Igarashi, I.
中科院分区：
文献类型：
--
作者：
Alhassan, A.;Iseki, H.;Igarashi, I.
在PCR基因扩增之前，快速、高效、可重复地分离DNA的程序对于诊断螺旋体至关重要。在本研究中，我们用预提取的DNA样本（QIAamp DNA Mini Kit和苯酚氯仿法）与FTA卡片上的血迹作为PCR模板，比较了PCR检测马伊勒菌的简易性和可靠性。虽然在比较的方法中观察到最小的检测限变化，但总体而言，使用预提取的DNA样本和在FTA卡片上发现的血液具有相当的检测限。这些结果表明，无需预先提取血样中的DNA，可以直接从FTA卡片上有效地检测马弓形虫感染。
Rapid, efficient, and reproducible procedures for isolating DNA before PCR gene amplification are essential for the diagnosis of piroplasms. In this study, we evaluated the ease and reliability of detecting Theileria equi by PCR using pre-extracted DNA samples ( by QIAamp DNA Mini Kit and phenolchloroform methods) compared with blood spotted on FTA cards as PCR templates. Although minimal variations in limit of detection were observed among the methods compared, overall, the use of pre-extracted DNA samples and blood spotted on FTA cards had comparable detection limits. These results indicate that T. equi infection can be efficiently detected directly from FTA cards by PCR without the need for preextraction of DNA from blood samples.