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从中国结膜蝇蛆病患者中采集的 Oestrus ovis(双翅目:Oestridae)幼虫的形态和分子鉴定。

Morphological and Molecular Identification of Oestrus ovis (Diptera: Oestridae) Larvae Collected from a Chinese Patient with Conjunctival Myiasis.

机构信息

Department of Pathogenic Biology and Immunology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, No.76 Yanta West Road, Xi'an, 710061, Shaanxi, China.

Zonglian Academy, Xi'an Jiaotong University Health Science Center, No. 76 Yanta West Road, Xi'an, 710061, Shaanxi, China.

出版信息

Acta Parasitol. 2022 Sep;67(3):1273-1281. doi: 10.1007/s11686-022-00573-x. Epub 2022 Jun 16.

Abstract

PURPOSE

Human conjunctival myiasis, which is often misdiagnosed or missed clinically, is commonly caused by Oestrus ovis larvae. Here, pathogenic identification was performed for two maggots collected from a patient from China, to provide a clinical scientific basis for diagnosis and treatment.

METHODS

Morphological identification was performed using a microscope. Oestrus mtDNA cox1 and rDNA 28S were selected as target genes for duplex PCR amplification, followed by cloning, sequencing, and identification.

RESULTS

Morphological examination showed that the maggots were approximately 1.0-1.5 mm long, long-oval-shaped, segmented, and covered with small spines, with a pair of hooks in the scolex and claw-like spines at the telson. Therefore, they were identified as the first-instar larvae of O. ovis. Duplex PCR detected products of approximately 400 and 200 bp, consistent with the size of designed cox1 and 28S D7a gene fragments, respectively. Sequences of cox1 and 28S D7a from the samples in question had 99.5-100.0% and 96.2-100.0% similarity (respectively) to GenBank sequences of O. ovis specimens known to parasitize sheep, goats, and humans. However, some 28S D7a sequences exhibited 89.7-90.6% similarity to GenBank sequences of Oestrus sp. known to parasitize Capra pyrenaica (Artiodactyla: Bovidae) (Iberian ibex). Therefore, we considered that the larvae infecting the patient originated from sheep or goats, not Iberian ibex. The phylogenetic trees supported this conclusion.

CONCLUSION

This study implemented the first duplex PCR molecular identification of O. ovis larvae parasitizing human eyes in China as a complementary approach to morphological identification. Our results indicate that molecular tools can be utilized to aid in the diagnosis of opthalmomyiasis.

摘要

目的

人眼蝇蛆病常被临床误诊或漏诊,多由羊狂蝇幼虫引起。本研究对 2 例来自中国患者眼部的蝇蛆进行致病性鉴定,为临床诊断和治疗提供科学依据。

方法

利用显微镜进行形态学鉴定,选择羊狂蝇 mtDNA cox1 和 rDNA 28S 为靶基因,进行双重 PCR 扩增、克隆、测序及鉴定。

结果

形态学检查发现,幼虫约 1.0-1.5mm 长,长椭圆形,分节,全身被小刺,头节前有一对钩,尾节有爪状刺,鉴定为羊狂蝇第 1 期幼虫。双重 PCR 检测到约 400bp 和 200bp 的产物,与 cox1 和 28S D7a 基因片段设计大小一致。所研究样本 cox1 和 28S D7a 序列与已知感染绵羊、山羊和人类的羊狂蝇 O. ovis 序列的相似性分别为 99.5-100.0%和 96.2-100.0%,但 28S D7a 序列与已知感染绵羊的羊狂蝇 Oestrus sp. 序列的相似性为 89.7-90.6%(偶蹄目:牛科)(伊比利亚羱羊)。因此,我们认为感染患者的幼虫来自绵羊或山羊,而不是伊比利亚羱羊。系统发育树支持这一结论。

结论

本研究首次对中国感染人眼的羊狂蝇幼虫进行双重 PCR 分子鉴定,为形态学鉴定提供了补充手段。研究结果表明,分子工具可用于辅助眼部蝇蛆病的诊断。

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