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巢式聚合酶链反应检测人疥螨细胞色素 c 氧化酶亚单位 1 基因诊断疥疮的效果。

The efficacy of a nested polymerase chain reaction in detecting the cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei var. hominis for diagnosing scabies.

机构信息

Department of Dermatology, Kangdong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea.

出版信息

Br J Dermatol. 2018 Oct;179(4):889-895. doi: 10.1111/bjd.16657. Epub 2018 Jul 13.

Abstract

BACKGROUND

A widespread scabies infestation, associated with long-term residence in nursing homes, is becoming an issue in high-income countries. Mineral oil examination is regarded as the gold standard in diagnosing scabies, but the sensitivity of this method is generally low - approximately 50%. Molecular techniques may contribute to enhancing the sensitivity of current tests for laboratory diagnosis of human scabies.

OBJECTIVES

To develop new primers for a nested polymerase chain reaction (PCR) for the cytochrome c oxidase subunit 1 (cox1) gene of Sarcoptes scabiei var. hominis to increase the sensitivity of a previously developed conventional PCR.

METHODS

Patients with clinically suspected scabies underwent dermoscopy-guided skin scraping with microscopic examination. The diagnosis was positive for scabies when mites or eggs were found under the microscope, and patients were then designated as 'microscopy positive'. Patients who presented with negative microscopic results were placed in the 'microscopy-negative' group. Skin scrapings were collected from both groups for PCR.

RESULTS

Of the total 63 samples, 28 were microscopy positive and 35 were negative with no differences in sex and age between the two groups. All microscopically proven cases of scabies were positive using the cox1 nested PCR. Among microscopy-negative samples, S. scabieiDNA was detected in nine. If sensitivity of the cox1 nested PCR is considered 100% [95% confidence interval (CI) 90·5-100], then sensitivity of microscopy is 75·7% (95% CI 58·8-88·2; P = 0·004).

CONCLUSIONS

Nested PCR can be successfully used as an alternative method for diagnosing suspected scabies. Therefore, infection control measures and treatments can be initiated before significant transmission occurs, minimizing the risk of outbreaks.

摘要

背景

一种广泛存在的疥疮感染与长期居住在养老院有关,这种情况在高收入国家正成为一个问题。矿物油检查被认为是诊断疥疮的金标准,但这种方法的敏感性通常较低 - 约为 50%。分子技术可能有助于提高目前用于实验室诊断人类疥疮的检测的敏感性。

目的

为了开发针对人疥螨 cox1 基因的嵌套聚合酶链反应(PCR)的新引物,以提高先前开发的常规 PCR 的敏感性。

方法

临床疑似疥疮的患者接受皮肤镜引导下的皮肤刮片和显微镜检查。当在显微镜下发现螨虫或卵时,诊断为疥疮阳性,然后将患者指定为“显微镜阳性”。呈现阴性显微镜结果的患者被归入“显微镜阴性”组。从两组患者中采集皮肤刮片进行 PCR。

结果

在总共 63 个样本中,28 个为显微镜阳性,35 个为阴性,两组患者的性别和年龄无差异。所有经显微镜证实的疥疮病例均使用 cox1 嵌套 PCR 呈阳性。在显微镜阴性样本中,检测到 9 例 S. scabieiDNA。如果 cox1 嵌套 PCR 的敏感性被认为是 100%[95%置信区间(CI)90.5-100],那么显微镜的敏感性为 75.7%(95% CI 58.8-88.2;P=0.004)。

结论

嵌套 PCR 可成功用作疑似疥疮的替代诊断方法。因此,可以在发生重大传播之前启动感染控制措施和治疗,最大限度地降低爆发的风险。

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