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首页> 外文期刊>The American Journal of Tropical Medicine and Hygiene >Paper chromatography hybridization: a rapid method for detection of Onchocerca volvulus DNA amplified by PCR.
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Paper chromatography hybridization: a rapid method for detection of Onchocerca volvulus DNA amplified by PCR.

机译：纸色谱杂交：一种检测PCR扩增的盘尾丝虫DNA的快速方法。

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Prior studies have shown that Onchocerca volvulus DNA can be detected in skin snips and in black flies after polymerase chain reaction (PCR) with primers specific for repeated "O-150" DNA sequences. We have adapted a paper chromatography hybridization assay (PCHA) to detect amplified O-150 DNA and compared this method to two established methods, namely agarose gel electrophoresis (AGE) and hybridization enzyme-linked immunosorbent assay (ELISA). The minimum amounts of purified O-150 DNA detected by PCHA, AGE, and ELISA were 5, 10, and 2 ng, respectively. The three methods had similar estimated sensitivities for detecting O. volvulus DNA amplified from skin snips from African subjects with onchocerciasis (88%, 84%, and 91%, respectively). No false positive results were observed with skin snips from uninfected control subjects. The paper chromatography hybridization assay detects PCR products in 30 minutes without electricity or special equipment. This technology brings DNA detection a step closer to widespread use in field settings.
机译：先前的研究表明，使用特异性针对重复“ O-150” DNA序列的引物，在聚合酶链反应（PCR）后，可在皮肤剪断和黑蝇中检测到盘尾线虫DNA。我们已采用纸色谱杂交测定法（PCHA）来检测扩增的O-150 DNA，并将该方法与两种已建立的方法进行比较，即琼脂糖凝胶电泳（AGE）和杂交酶联免疫吸附测定（ELISA）。通过PCHA，AGE和ELISA检测到的纯化O-150 DNA的最小量分别为5、10和2 ng。这三种方法在检测从盘尾丝虫病的非洲受试者的皮肤剪中扩增出的肠弯曲杆菌DNA时，估计灵敏度相似（分别为88％，84％和91％）。未感染对照受试者的皮肤剪没有观察到假阳性结果。纸色谱杂交测定法可在30分钟内无需电力或专用设备即可检测PCR产物。这项技术使DNA检测更加接近在现场环境中的广泛使用。

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