Early Detection of Histomoniasis in Blood Samples by PCR and Sequencing.

Vijay Durairaj, Emily Barber, Ryan Vander Veen
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Abstract

Histomoniasis is a deadly disease of turkeys causing devastating economic losses to the poultry industry. In field outbreaks, a presumptive diagnosis is made based on gross pathology lesions and confirmed by histopathology. An early detection tool with quick turnaround time is needed to prevent the spread of histomoniasis. With this objective, two studies were conducted in turkeys. In Study 1, 40 poults were housed in two pens (20 poults/pen) and challenged at 14 days of age with Histomonas meleagridis by intracloacal route. Blood samples were collected 4 days postchallenge. Fifty-five percent (22/40) of the blood samples tested positive for H. meleagridis based on PCR using primers targeted against the 18S rRNA gene and confirmed by sequencing. In Study 2, 40 poults were housed in two groups and raised in floor pens. Groups 1 and 2 served as negative and challenge controls, respectively. At 14 days of age, the birds in Group 2 were challenged with H. meleagridis by intracloacal route. Blood samples were collected 2 days postchallenge. Five percent (1/20) of the blood samples tested positive for H. meleagridis, based on PCR and confirmed by sequencing. The results from both studies indicate that H. meleagridis DNA can be detected in the blood samples by PCR and confirmed by sequencing as early as 4 days postchallenge. This early detection method could be applied in field outbreaks to detect and confirm histomoniasis as early as possible.

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利用 PCR 和测序技术早期检测血液样本中的组织滴虫病。
组织单胞菌病是火鸡的一种致命疾病,给家禽业造成了毁灭性的经济损失。在田间疫情爆发时,可根据大体病理学病变做出推定诊断,并通过组织病理学进行确诊。为防止组织单胞菌病的传播,需要一种可快速周转的早期检测工具。为此,我们在火鸡身上进行了两项研究。在研究 1 中,40 只火鸡被饲养在两个围栏中(每个围栏 20 只),并在 14 日龄时通过球内途径感染组织单胞菌。挑战后 4 天采集血液样本。使用针对 18S rRNA 基因的引物进行 PCR 检测,55%(22/40)的血液样本对 H. meleagridis 检测呈阳性,并通过测序得到证实。在研究 2 中,40 只小鸡被分成两组,在地栏中饲养。第 1 组和第 2 组分别作为阴性对照组和阳性对照组。14 日龄时,第 2 组的家禽通过球内途径接受 H. meleagridis 的挑战。挑战后 2 天采集血液样本。根据聚合酶链式反应(PCR),5%(1/20)的血液样本对 H. meleagridis 检测呈阳性,并经测序证实。这两项研究的结果表明,通过聚合酶链式反应(PCR)可以检测到血液样本中的 H. meleagridis DNA,并可在感染后 4 天内通过测序确认。这种早期检测方法可用于野外爆发的组织单胞菌病的早期检测和确诊。
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