畜牧兽医杂志2025，Vol.44Issue(6)：50-54,62,6.DOI:10.7606/j.issn.1004-6704.2025.06.011
基于猫STK40基因的弓形虫感染新型PCR检测方法的建立与评价
Development and Evaluation of a Novel PCR Assay for Toxoplasma gondii Infection Based on Feline STK40 Gene
摘要
Abstract
This study aimed to establish a novel PCR detection method based on the feline host STK40 gene to improve the diagnostic accuracy and efficiency for Toxoplasma gondii infection.Transcriptome analysis revealed that the STK40 gene was significantly downregulated after T.gondii infection,and no homologous gene exists in the T.gondii genome,making it an ideal target for specific detection.Primers targeting the variable region of the STK40 gene(amplicon length:207 bp)were designed,and optimal PCR conditions(annealing temperature:55.6 ℃,time:30 s)were determined through gradient optimization.Sensitivity tests demonstrated effective amplification at DNA concentrations as low as 8.819 and 5.745 fg/μL.Specificity assays confirmed positive results only for T.gondii-infected feline DNA,with no cross-reactivity to other parasites.Re-peatability tests indicated high stability and reliability,with a positive detection rate of 40％(4/10)in practical applica-tions.The established STK40-based PCR assay exhibited high sensitivity and specificity under laboratory conditions,though its clinical applicability requires further validation with larger sample sizes.This study provides a reliable tool for the early diagnosis of T.gondii infection,particularly ad-dressing the prevention and control needs of high-risk popu-lations such as pregnant women and immunocompromised individuals,highlighting its significant clinical and public health.关键词
弓形虫/猫/STK40基因/PCR检测/分子诊断Key words
Toxoplasma gondii/feline/STK40 gene/PCR detection/molecular diagnosis分类
畜牧业引用本文复制引用
翟斌涛,李甲,包碧波,马少霞,谢世臣,周雅馨,王玮玮,张继瑜..基于猫STK40基因的弓形虫感染新型PCR检测方法的建立与评价[J].畜牧兽医杂志,2025,44(6):50-54,62,6.基金项目
兰州市青年科技人才创新项目(2023-QN-73) （2023-QN-73）
陇原青年创新创业人才项目(2024QNTD41) （2024QNTD41）
甘肃省自然基金(23JRRA562) （23JRRA562）
国家肉牛牦牛产业技术体系项目(CARS-37) （CARS-37）
