Animal Parasitic Diseases Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, USDA, Beltsville, MD 20705,
Animal Parasitic Diseases Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, USDA, Beltsville, MD 20705.
Avian Dis. 2022 Jun;66(2):181-185. doi: 10.1637/aviandiseases-D-22-00001. Epub 2022 Jul 11.
The purpose of this study was to compare micro-oocyst counts of to PCR analysis of intestinal DNA from smears of duodenum, jejunum/ileum, and cecum of chickens infected with , , or oocysts. Broiler chicks were infected in triplicate with various doses of , , or oocysts and were necropsied 5-6 days later to recover duodenal, jejunal, or cecal tissue for micro-oocyst count and for DNA recovery. Micro-oocyst counts were done independently by three individuals. Micro-oocyst counts and PCR directed to ITS1 rDNA or to a single-copy orthologue (SCO 5995) displayed a linear relationship with oocyst dose for each species. A strong correlation was found between mean micro-oocyst counts and both PCR assays for ( = 0.78-0.94), ( = 0.79-0.91), and ( = 0.85-0.96). There was good agreement between ITS1 and SCO 5995 PCR assays: ( = 0.92), ( = 0.79), and ( = 0.93). However, only ITS1 PCR analysis corroborated micro-oocyst counts of oocyst DNA recovered from -infected broiler chickens submitted to a poultry diagnostic laboratory. These findings suggest that ITS1 PCR or SCO PCR can validate traditional micro-oocyst counts used in quantifying infection in chickens. Additional studies may provide a method for estimating the relative abundance of each species in a natural infection.
本研究旨在比较感染 、 或 卵囊的鸡的十二指肠、空肠/回肠和盲肠涂片的肠道 DNA 的 PCR 分析与微卵囊计数。将肉鸡重复感染各种剂量的 、 或 卵囊，并在 5-6 天后剖检以回收十二指肠、空肠或盲肠组织进行微卵囊计数和 DNA 回收。微卵囊计数由三个人独立进行。微卵囊计数和针对 ITS1 rDNA 或单拷贝同源物（SCO 5995）的 PCR 与每种 物种的卵囊剂量呈线性关系。发现 （= 0.78-0.94）、 （= 0.79-0.91）和 （= 0.85-0.96）的平均微卵囊计数与两种 PCR 检测之间存在很强的相关性。ITS1 和 SCO 5995 PCR 检测之间具有良好的一致性： （= 0.92）、 （= 0.79）和 （= 0.93）。然而，只有 ITS1 PCR 分析证实了从提交给家禽诊断实验室的感染 - 的肉鸡中回收的卵囊 DNA 的微卵囊计数。这些发现表明，ITS1 PCR 或 SCO PCR 可以验证用于量化鸡中 感染的传统微卵囊计数。进一步的研究可能提供一种方法来估计自然感染中每种 物种的相对丰度。
