猫泛白细胞减少症病毒纳米PCR检测方法的建立与初步应用

Establishment and preliminary application of a Nano-PCR method for detection of feline panleukopenia virus

为建立一种灵敏且快速检测猫泛白细胞减少症病毒(FPV)的分子检测方法,本研究采用纳米颗粒与普通PCR技术相结合,通过使用Oligo 7设计FPV的特异性引物,建立FPV Nano-PCR分子检测方法,并与普通PCR方法进行对比分析。结果显示,所建立的Nano-PCR的敏感度是普通PCR的10倍(10 fg/μL);特异性试验表明,Nano-PCR只检测猫泛白细胞减少症病毒时呈阳性反应,而检测猫杯状病毒(FCV)、猫疱疹病毒(FHV)以及F81细胞cDNA/DNA时呈阴性反应。利用两种方法对采集的26份临床病料进行平行检测,发现Nano-PCR与普通PCR阳性率分别为53.84%和42.31%。上述结果表明,所建方法具有敏感性好、特异强、简捷快速等优点,可应用于临床样品的快速检测和鉴定,为猫泛白细胞减少症的早期诊断和防控提供了一定技术保障。

In order to establish a sensitive and rapid molecular detection method for feline panleukopenia virus(FPV).In this study,nanoparticles were combined with conventional PCR technology,and specific primers for FPV were designed by using Oligo 7 to establish a molecular detection method for FPV Nano-PCR,which was compared with conventional PCR method for analysis.The results showed that the sensitivity of Nano-PCR was 10 times that of conventional PCR(10 fg/μL);Specificity test showed that NanoPCR was positive only for feline panleukopenia virus,but negative for feline calicivirus(FCV),feline herpes virus(FHV) and F81 cell c DNA/DNA.The results showed that the positive rate of Nano-PCR and conventional PCR was 53.84% and 42.31%,respectively.These results indicated that the established method has the advantages of good sensitivity,specificity,simplicity and rapidness,which can be applied to the rapid detection and identification of clinical samples,providing a technical guarantee for the early diagnosis,prevention and control of feline panleukopenia.