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粪类圆线虫的诊断:尿液中寄生虫衍生DNA的检测。

Diagnosis of Strongyloides stercoralis: Detection of parasite-derived DNA in urine.

作者信息

Lodh Nilanjan, Caro Reynaldo, Sofer Shterna, Scott Alan, Krolewiecki Alejandro, Shiff Clive

机构信息

Marquette University, College of Health Sciences, Department of Clinical Laboratory Science, 561, North 15 Street, Milwaukee, WI 53233, USA.

Instituto de Investigaciones en Enfermedades Tropicales. Universidad Nacional de Salta, sede regional Oran, Alvarado 751, Orán (4530), Salta, Argentina.

出版信息

Acta Trop. 2016 Nov;163:9-13. doi: 10.1016/j.actatropica.2016.07.014. Epub 2016 Jul 22.

Abstract

Detecting infections of Strongyloides stercoralis is arduous and has low sensitivity. Clinically this is a major problem because chronic infections may disseminate in the host and lead to a life threatening condition. Epidemiologically, S. stercoralis is often missed in surveys as it is difficult to identify by standard stool examination procedures. We present, for the first time, evidence that the infection can be detected in filtered urine samples collected and processed in the field and subsequently assayed for the presence of parasite DNA. Urine specimens (∼40mL) were collected from 125 test and control individuals living in rural and peri-urban regions of Northern Argentina. From the same individuals, fresh stool specimens were processed using three different copropological methods. Urine specimens were filtered in the field through a 12.5cm Whatman No. 3 filter. The filters were dried and packed individually in sealable plastic bags with desiccant and shipped to a laboratory where DNA was recovered from the filter and PCR-amplified with primers specific to a dispersed repetitive sequence. Prevalence of S. stercoralis infection by stool culture and direct examination was 35/125 (28%), In contrast, PCR-based detection of parasite-specific trans-renal DNA in urine indicated that 56/125 (44.8%) carried the parasite. Of the patients that tested positive for urine-based parasite DNA, approximately half also tested positive in their stool specimens. There were 6.4% of cases where parasite larvae were seen in the stool but no DNA was amplified from the urine. As proof of principle, DNA amplification from urine residue reveals significantly more cases of S. stercoralis infection than the current standard stool examination techniques. Additional work is required to establish the relative utility, sensitivity and specificity of urine-based analysis compared to parasitological and nucleic acid detection from stool for clinical and epidemiological detection for S. stercoralis infection.

摘要

检测粪类圆线虫感染既艰巨又灵敏度低。临床上这是个大问题,因为慢性感染可能在宿主体内播散并导致危及生命的状况。在流行病学方面,粪类圆线虫在调查中常常被漏检,因为通过标准粪便检查程序很难识别。我们首次提供证据表明,在野外收集和处理的过滤尿液样本中可以检测到这种感染,随后对寄生虫DNA的存在进行检测。从居住在阿根廷北部农村和城郊地区的125名检测对象和对照个体中收集尿液样本(约40毫升)。从同一批个体中,使用三种不同的粪便学方法处理新鲜粪便样本。尿液样本在野外通过一个12.5厘米的沃特曼3号滤纸过滤。滤纸干燥后单独装在带有干燥剂的可密封塑料袋中,然后运至实验室,从滤纸上提取DNA并用针对分散重复序列的引物进行PCR扩增。通过粪便培养和直接检查,粪类圆线虫感染的患病率为35/125(28%)。相比之下,基于PCR检测尿液中寄生虫特异性经肾DNA表明,56/125(44.8%)携带该寄生虫。在尿液中寄生虫DNA检测呈阳性的患者中,约一半在粪便样本检测中也呈阳性。有6.4%的病例在粪便中发现了寄生虫幼虫,但尿液中未扩增出DNA。作为原理验证,从尿液残渣中进行DNA扩增显示,粪类圆线虫感染病例比当前标准粪便检查技术显著更多。与粪便寄生虫学和核酸检测相比,还需要进一步开展工作来确定基于尿液分析在粪类圆线虫感染临床和流行病学检测中的相对效用、灵敏度和特异性。

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