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猫体内 属的检测:通过聚合酶链式反应(PCR)和高分辨率熔解曲线分析(HRM)对鼻拭子、口腔拭子和结膜拭子进行分析。 (注:原文中“spp.”处信息缺失,翻译时保留原样)

Detection of spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM.

作者信息

Alves-Martin Maria Fernanda, Bertozzo Thainá Valente, Aires Isabella Neves, Manzini Suzane, Paixão-Marques Mirian Dos Santos, Guiraldi Lívia Maísa, Dos Santos Wesley José, Sánchez Gabriela Pacheco, Curci Vera Cláudia Lorenzetti Magalhães, Richini-Pereira Virgínia Bodelão, Lucheis Simone Baldini

机构信息

Department of Biology and Animal Science, School of Engineering, Sao Paulo State University (UNESP), Ilha Solteira 15385-000, SP, Brazil.

Department of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, Brazil.

出版信息

Animals (Basel). 2023 Jul 31;13(15):2468. doi: 10.3390/ani13152468.

Abstract

BACKGROUND AND OBJECTIVES

Feline leishmaniasis (FeL) is caused by several species of parasites of the genus . The disease can occur with the presence or absence of clinical signs, similar to those observed in other common infectious diseases. In endemic regions for FeL, the infection has been associated with dermatological lesions. Therefore, considering the search for less invasive and more effective diagnostic techniques, we aimed to investigate the presence of spp. in domestic cats through Polymerase Chain Reaction (PCR) and high-resolution melting (HRM) analyses of conjunctival, oral, and nasal epithelial cells, and we detected the presence of anti- IgG antibodies from serological techniques of the Immunofluorescent Antibody Test (IFAT) and ELISA.

METHODS

The PCR and HRM for detection of spp. were performed on 36 samples of epithelial cells from the conjunctiva of male and female cats, collected using sterile swabs. The serological tests IFAT and ELISA were also performed.

RESULTS

The prevalence of infection was 11.1% (4/36) by PCR assay, and those results were confirmed for species using the HRM technique. Twenty-four cats (24/36 = 66.7%) were reactive to the IFAT and twenty-two cats were reactive by the ELISA technique (22/36 = 61.1%).

INTERPRETATION AND CONCLUSIONS

The use of conjunctival swabs was shown to be a non-invasive, practical, and easy-to-perform technique, and in addition to the genetic sequencing and HRM, it was able to identify the parasitic DNA of in cats. This technique can be used for screening diagnosis in future epidemiological surveys of FeL and can be used as a complement to clinical and/or serological tests, as well as associating the clinical history of the animal, for the diagnostic conclusion.

摘要

背景与目的

猫利什曼病(FeL)由利什曼原虫属的几种寄生虫引起。该疾病可能出现或不出现临床症状,与其他常见传染病中观察到的症状相似。在FeL的流行地区,感染与皮肤病变有关。因此,考虑到寻求侵入性较小且更有效的诊断技术,我们旨在通过聚合酶链反应(PCR)和结膜、口腔及鼻上皮细胞的高分辨率熔解(HRM)分析,研究家猫中利什曼原虫属物种的存在情况,并通过免疫荧光抗体试验(IFAT)和酶联免疫吸附测定(ELISA)的血清学技术检测抗利什曼原虫IgG抗体的存在。

方法

使用无菌拭子收集36份雄性和雌性猫结膜上皮细胞样本,进行检测利什曼原虫属物种的PCR和HRM。还进行了血清学检测IFAT和ELISA。

结果

通过PCR检测,利什曼原虫感染率为11.1%(4/36),使用HRM技术对这些结果进行了物种确认。24只猫(24/36 = 66.7%)对IFAT呈阳性反应,22只猫通过ELISA技术呈阳性反应(22/36 = 61.1%)。

解读与结论

使用结膜拭子被证明是一种非侵入性、实用且易于操作的技术,除了基因测序和HRM外,它能够识别猫体内利什曼原虫的寄生DNA。该技术可用于未来FeL流行病学调查的筛查诊断,并可作为临床和/或血清学检测的补充,以及结合动物的临床病史,用于诊断结论。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1173/10417458/f6849f58e3b6/animals-13-02468-g001.jpg

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