Ulucesme Mehmet Can, Ozubek Sezayi, Aktas Munir
Department of Parasitology, Faculty of Veterinary Medicine, University of Fırat, Elazığ 23200, Türkiye.
Vet Sci. 2024 Oct 1;11(10):466. doi: 10.3390/vetsci11100466.
We developed and evaluated a semi-nested PCR assay for the detection of infection in goats based on the sequence of the gene. Following in silico screening, the specificity of the primers was assessed using reference DNA samples, including , , , , , , , and . To determine the sensitivity of the method, blood infected with 2% parasitemia of was diluted to 10-fold serial dilutions. The method specifically amplified a 438 bp fragment of DNA, but did not demonstrate cross-amplification with the other hemoparasites tested. The sensitivity assay indicated that this PCR method was able to detect infection at a dilution of 10 of 2% parasitemia (0.074 parasites/200 µL). Ninety-seven blood samples collected from goats were used to analyze for , and the infection was detected in 18.5% of the goats. Additionally, the method was also applied to 44 field DNA samples that were detected to be positive for by reverse line blotting (RLB), and showed 84.1% agreement. The findings revealed that newly developed semi-nested PCR can detect infections in goats with high sensitivity and specificity.
我们基于该基因序列开发并评估了一种用于检测山羊感染的半巢式聚合酶链反应（PCR）检测方法。经过计算机筛选后，使用包括……（此处原文缺失具体参考DNA样本名称）等参考DNA样本评估引物的特异性。为确定该方法的灵敏度，将感染2%疟原虫血症的血液进行10倍系列稀释。该方法特异性地扩增出一段438 bp的该DNA片段，但与所检测的其他血液寄生虫未出现交叉扩增。灵敏度检测表明，这种PCR方法能够在2%疟原虫血症稀释至10⁻⁶时检测到感染（0.074个寄生虫/200微升）。从山羊采集的97份血液样本用于分析该感染，在18.5%的山羊中检测到感染。此外，该方法还应用于44份通过反向线印迹法（RLB）检测为该感染阳性的野外DNA样本，显示出84.1%的一致性。研究结果表明，新开发的半巢式PCR能够以高灵敏度和特异性检测山羊中的该感染。
