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比较多重 PCR 与粪检后回收卵 PCR 检测犬细粒棘球蚴和带绦虫感染。

Comparison of multiplex copro PCR with coproscopy followed by PCR on recovered eggs for the detection of Echinococcus granulosus and Taenia spp. infection in dogs.

机构信息

Division of Parasitology, Kimron Veterinary Institute, P.O.B. 12, Bet Dagan 50250, Israel.

Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, Rehovot, Israel.

出版信息

Vet Parasitol. 2023 Mar;315:109885. doi: 10.1016/j.vetpar.2023.109885. Epub 2023 Jan 20.

Abstract

Echinococcosis and taeniasis are important helminth diseases that carry considerable impact on human and animal health. Domestic dogs and other canids are definitive hosts for several parasites of this group, including Echinococcus granulosus, Taenia multiceps, T. ovis, T. hydatigena and E. multilocularis. Detection of infection in dog populations is imperative for estimating the risk to susceptible humans and animals, and for its mitigation through prevention measures in dogs, other animals and humans. To date, identification of taeniid eggs, antigens or DNA in fecal samples are the most practical diagnostic modalities available for canine definitive hosts. Although widely used for this purpose, there is limited information comparing copro PCR and combined coproscopy-PCR protocols for the detection of taeniids. In the current study, a widely used multiplex PCR was performed on a large number of dog fecal samples using DNA extracted directly from feces. The samples were also tested by fecal flotation and coproscopy, eggs were isolated from microscopically-positive samples and extracted DNA was tested using the same multiplex PCR. The total number of taeniid positive samples detected using both methods was 46/317 (14.5%), including 10/317 (3.2%) E. granulosus positive samples. Both copro PCR and coproscopy have identified an equal number of samples as taeniid positive (n = 32). However, for the purpose of identification to species level, the copro PCR was significantly more sensitive than coproscopy followed by PCR on isolated eggs (sensitivity 0.7 vs. 0.41, p = 0.012), with 32/317 (10.1%) and 19/317 (6%) positive samples identified, respectively. The difference in identification of E. granulosus was highly apparent, as the majority of the E. granulosus positive samples (8/10) were detected by the copro PCR only. Coproscopy and egg PCR have identified 5/317 (1.6%) positive samples not detected by the copro PCR, including only a single sample (0.3%) positive for E. granulosus. Adding these positive samples to those identified by the copro PCR did not significantly improve the overall sensitivity (p = 0.074). Therefore, using both copro PCR and coproscopy in parallel may not be advantageous for taeniid detection and identification, at least until the egg PCR is further optimized and performs better. These results should be weighed against the different advantages that coproscopy based approach may offer.

摘要

包虫病和带绦虫病是重要的寄生虫病,对人类和动物健康有重大影响。家养狗和其他犬科动物是该组几种寄生虫的终末宿主,包括细粒棘球绦虫、多头绦虫、绵羊绦虫、细粒棘球绦虫和泡状带绦虫。检测犬群中的感染情况对于评估易感人群和动物的风险以及通过在犬、其他动物和人类中采取预防措施来减轻风险至关重要。迄今为止,在粪便样本中检测带绦虫卵、抗原或 DNA 是最实用的犬科终末宿主诊断方法。尽管广泛用于此目的,但比较 copro PCR 和联合 coproscopy-PCR 检测带绦虫的协议的信息有限。在本研究中,使用直接从粪便中提取的 DNA 对大量犬粪便样本进行了广泛使用的多重 PCR。还通过粪便漂浮和 coproscopy 对样本进行了检测,从显微镜阳性样本中分离出虫卵,并使用相同的多重 PCR 对提取的 DNA 进行了检测。使用这两种方法检测到的总带绦虫阳性样本数为 46/317(14.5%),其中包括 10/317(3.2%)的细粒棘球绦虫阳性样本。copro PCR 和 coproscopy 均鉴定出相同数量的带绦虫阳性样本(n=32)。然而,为了鉴定到种水平,copro PCR 明显比随后的分离卵 PCR 更敏感(灵敏度 0.7 比 0.41,p=0.012),分别鉴定出 32/317(10.1%)和 19/317(6%)阳性样本。细粒棘球绦虫的鉴定差异非常明显,因为大多数细粒棘球绦虫阳性样本(8/10)仅通过 copro PCR 检测到。coscopy 和卵 PCR 鉴定出 copro PCR 未检测到的 5/317(1.6%)阳性样本,其中仅 1 个样本(0.3%)为细粒棘球绦虫阳性。将这些阳性样本添加到 copro PCR 鉴定的阳性样本中并不会显著提高总体敏感性(p=0.074)。因此,在卵 PCR 进一步优化并表现更好之前,平行使用 copro PCR 和 coproscopy 可能对带绦虫检测和鉴定没有优势。应该权衡基于 coproscopy 的方法可能提供的不同优势。

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