■当通过健康监测使用具有保证的微生物学和遗传质量的实验动物进行动物实验时,可以通过可靠和可重复的实验结果来提高研究质量。因此,健康监测需要快速准确地诊断实验动物的传染病。
■本研究提出了一种用于快速检测12种传染性病原体的市售多重实时PCR(mRT-PCR)测定法的性能评估(第1组:仙台病毒[SeV,正式的鼠呼吸道病毒],支原体属。,嗜肺分枝杆菌,和Heylii啮齿杆菌;第2组:螺杆菌属。,鼠诺如病毒[MNV],鼠肝炎病毒[MHV],和沙门氏菌。;组3:金黄色葡萄球菌,念珠链球菌,Kutscheri棒杆菌,和铜绿假单胞菌)。为了评估mRT-PCR测定的功效,分析了102个包含粪便和盲肠标本的临床样品。然后将所得数据与来自序列分析的结果进行比较以进行验证。
■测定的检出限范围为每个反应1至100个拷贝。涉及各种病毒和细菌的特异性测试表明菌株之间没有交叉反应性。此外,该测定表现出良好的重现性,测定间和测定内变异的平均变异系数低于3%。总阳性率为52.9%(n=54),mRT-PCR检测结果与序列分析结果匹配(κ=1)。MHV(n=29,28.4%)是最常见的病原体,其次是螺杆菌。(n=28,27.5%),R.heylii(n=18,17.6%),支原体属。(n=14,13.7%),MNV(n=12,11.8%),金黄色葡萄球菌(n=9,8.8%),铜绿假单胞菌(n=4,3.9%),和肺炎链球菌(n=1,0.9%)。
■该测定提供了100分钟的快速周转时间,包括30分钟用于DNA制备和70分钟用于靶DNA/RNA扩增。它确保准确性,尽量减少假阳性或阴性,使其成为同时检测许多样品中传染病的便捷工具。总的来说,提议的-d测定法有望有效检测实验动物健康监测中最重要的病原体。
UNASSIGNED: Research quality can be improved with reliable and reproducible experimental results when animal experiments are conducted using laboratory animals with guaranteed microbiological and genetic quality through health monitoring. Therefore, health monitoring requires the rapid and accurate diagnosis of infectious diseases in laboratory animals.
UNASSIGNED: This study presents a performance evaluation of a commercially available multiplex real-time PCR (mRT-PCR) assay for the rapid detection of 12 infectious pathogens (Set 1: Sendai virus [SeV, formally murine respirovirus], Mycoplasma spp., Rodentibacter pneumotropicus, and Rodentibacter heylii; Set 2: Helicobacter spp., Murine norovirus [MNV], Murine hepatitis virus [MHV], and Salmonella spp.; Set 3: Staphylococcus aureus, Streptobacillus moniliformis, Corynebacterium kutscheri, and Pseudomonas aeruginosa). To evaluate the efficacy of the mRT-PCR assay, 102 clinical samples encompassing fecal and cecal specimens were analyzed. The resulting data were then compared with the findings from sequence analysis for validation.
UNASSIGNED: The assay\'s detection limit ranged from 1 to 100 copies per reaction. Specificity testing involving various viruses and bacteria indicated no cross-reactivity between strains. Additionally, the assay exhibited good reproducibility, with mean coefficients of variation for inter- and intra assay variation below 3%. The overall positive rate was 52.9% (n = 54), with the mRT-PCR assay findings matching sequence analysis results (κ = 1). MHV (n = 29, 28.4%) was the most prevalent pathogen, followed by Helicobacter spp. (n = 28, 27.5%), R. heylii (n = 18, 17.6%), Mycoplasma spp. (n = 14, 13.7%), MNV (n = 12, 11.8%), S. aureus (n = 9, 8.8%), P. aeruginosa (n = 4, 3.9%), and R. pneumotropicus (n = 1, 0.9%).
UNASSIGNED: This assay offers a rapid turnaround time of 100 min, including 30 min for DNA preparation and 70 min for target DNA/RNA amplification. It ensures accuracy, minimizing false positives or negatives, making it a convenient tool for the simultaneous detection of infectious diseases in many samples. Overall, the propose‑d assay holds promise for the effective detection of the most important pathogens in laboratory animal health monitoring.