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一种用于检测海狗肺线虫粪便和鼻拭子的新型定量实时PCR诊断检测方法。

A novel quantitative real-time PCR diagnostic assay for fecal and nasal swab detection of an otariid lungworm, .

作者信息

Williams Kalani M, Fessler M K, Bloomfield R A, Sandke William D, Malekshahi Clara R, Keroack Caroline D, Duignan Pádraig J, Torquato Samantha D, Williams Steven A

机构信息

Smith College, Department of Biological Sciences, Northampton, MA, 01063, USA.

The Marine Mammal Center, Sausalito, CA, 94965, USA.

出版信息

Int J Parasitol Parasites Wildl. 2020 May 18;12:85-92. doi: 10.1016/j.ijppaw.2020.04.012. eCollection 2020 Aug.

Abstract

also known as sea lion lungworm, is a metastrongyloid nematode that infects otariid hosts, such as the charismatic California sea lion, causes bronchointerstitial pneumonia, respiratory distress, reduced ability to swim, dive and hunt and as a result, increased mortality particularly in young animals. Respiratory disease is a leading cause of stranding and admission to rehabilitation centers on the Pacific coast. Low-coverage genomic sequencing of four individuals analyzed through Galaxy's RepeatExplorer identified a novel repeat DNA family we employed to design a sensitive quantitative PCR (qPCR) assay for diagnosing infections from fecal or sputum samples. The assay detects as little as 10 fg of DNA and a linear regression model developed using a standard curve can be used to estimate the concentration of DNA in a sample, ± 0.015 ng. This knowledge can be leveraged to estimate the level of parasite burden, which can be used to design improved treatments for animals in rehabilitation. Improved treatment of infections will aid in more animals being successfully released back into the wild.

摘要

也被称为海狮肺线虫,是一种后圆线虫类线虫,感染海兽宿主,如具有魅力的加利福尼亚海狮,会导致支气管间质性肺炎、呼吸窘迫、游泳、潜水和捕猎能力下降,结果导致死亡率增加,尤其是在幼兽中。呼吸系统疾病是太平洋海岸动物搁浅和进入康复中心的主要原因。通过Galaxy的RepeatExplorer对四个个体进行的低覆盖基因组测序鉴定出一个新的重复DNA家族,我们利用该家族设计了一种灵敏的定量PCR(qPCR)检测方法,用于诊断粪便或痰液样本中的感染。该检测方法能检测低至10 fg的DNA,使用标准曲线建立的线性回归模型可用于估计样本中DNA的浓度,误差为±0.015 ng。这些知识可用于估计寄生虫负荷水平,进而用于为康复中的动物设计更好的治疗方法。改善感染治疗将有助于更多动物成功放归野外。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48f9/7256429/b71fa98ae989/fx1.jpg

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