Accurate diagnosis of foetal sex in pregnant mare is helpful for many breeders, both for private or commercial purposes. In this study, in order to pre‐natal foetal sexing in equine, we used TaqMan duplex real‐time PCR to detect the specific regions of SRY and TSPY genes on extracted cell‐free foetal DNA from maternal blood. Peripheral blood samples from 50 pregnant Arabian mares with singleton foetuses were collected. Cell‐free foetal DNA was extracted from maternal plasma, and duplex real‐time PCR assays were performed with TaqMan probes and primers. Amplification of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) gene was used as control of DNA extraction procedure. From the 50 sampled mares, 28 cases had female and 22 mares had male foetuses. The final results for 46 samples were conclusive, and from them, 43 cases were predicted correctly. Sensitivity, specificity and accuracy of the test were 90.48%, 96% and 93.48%, respectively. In conclusion, a TaqMan duplex real‐time PCR was set up to pre‐natal detection of foetal sex in equine. The method was fast and decreased the false‐positive and false‐negative results. The technique can be used as a routine procedure in farms by collecting only a blood sample.

中文翻译：

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TaqMan双重实时PCR测定法对马中胎儿性别的非侵入性产前评估的效率评估

准确诊断怀孕母马的胎儿性别对于许多育种者都是有用的，无论是出于私人目的还是出于商业目的。在这项研究中，为了在马中进行产前胎儿性别鉴定，我们使用TaqMan双工实时PCR检测从母体血液中提取的无细胞胎儿DNA上SRY和TSPY基因的特定区域。收集了来自50具单胎胎儿的阿拉伯母马的外周血样本。从母体血浆中提取无细胞胎儿DNA，并使用TaqMan探针和引物进行双重实时PCR分析。使用3-磷酸甘油醛脱氢酶（GAPDH）基因的扩增作为DNA提取程序的对照。在50个母马中，有28例为雌性，22例为雄性。46份样本的最终结果是结论性的，并据此正确预测了43例。测试的灵敏度，特异性和准确性分别为90.48％，96％和93.48％。总之，已建立TaqMan双工实时PCR来对马中的胎儿性别进行产前检测。该方法快速，减少了假阳性和假阴性结果。通过仅收集血液样本，该技术可以用作农场的常规程序。