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一种新型、高敏感的核酸扩增检测方法用于利什曼病的诊断及其对循环游离 DNA 的检测。

A Novel, Highly Sensitive Nucleic Acid Amplification Test Assay for the Diagnosis of Loiasis and its Use for Detection of Circulating Cell-Free DNA.

机构信息

Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.

Genomics Unit, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.

出版信息

J Infect Dis. 2023 Oct 3;228(7):936-943. doi: 10.1093/infdis/jiad186.

Abstract

Mass drug administration programs targeting filarial infections depend on diagnostic tools that are sensitive and specific. The coendemicity of Loa loa with other filarial species often hampers the control programs. LL2634 was identified as the most promising target among several highly repeated targets, with sensitivity between 500 ag and 1 fg of genomic DNA. Using DNA from infected individuals, LL2643 quantitative polymerase chain reaction (qPCR) was positive in all individuals. LL2643 was detected in plasma-derived circulating cell-free DNA (ccfDNA) from 48 of 53 microfilariae-positive patients. Detection of ccfDNA in urine was possible, but it occurred rarely among those tested. Importantly, LL2643 ccfDNA became undetectable within 1 month following diethylcarbamazine (DEC) treatment and remained negative for at least a year. LL2643 offers a more sensitive and specific target for detection of L. loa infection and would be easily configurable to a point-of-contact assay. Clinical Trials Registration. NCT00001230 and NCT00090662.

摘要

大规模药物治疗项目针对的是丝虫感染,需要依靠敏感且特异的诊断工具。盘尾丝虫与其他丝虫种的共感染常常会给防治项目带来阻碍。在多个高度重复的靶标中,LL2634 被确定为最有希望的靶标,其对基因组 DNA 的检测灵敏度在 500 ag 到 1 fg 之间。利用感染者的 DNA,所有感染者的 LL2643 定量聚合酶链反应(qPCR)均为阳性。在 53 名微丝蚴阳性患者中,有 48 人从血浆衍生的循环无细胞 DNA(ccfDNA)中检测到了 LL2643。在尿液中也可以检测到 ccfDNA,但在检测到的样本中很少出现。重要的是,在乙胺嗪(DEC)治疗后 1 个月内,LL2643 的 ccfDNA 检测不到,并且至少在一年内保持阴性。LL2643 为检测盘尾丝虫感染提供了一个更敏感和特异的靶标,并且易于配置成接触点检测。临床试验注册。NCT00001230 和 NCT00090662。

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The art of obtaining a high yield of cell-free DNA from urine.从尿液中获取高产量无细胞 DNA 的技术。
PLoS One. 2020 Apr 6;15(4):e0231058. doi: 10.1371/journal.pone.0231058. eCollection 2020.

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