Calvert Amanda E, Biggerstaff Brad J, Tanner Nathan A, Lauterbach Molly, Lanciotti Robert S
Arboviral Diseases Branch, Division of Vector-Borne Diseases, U.S. Centers for Disease Control and Prevention, Fort Collins, CO, United States of America.
New England Biolabs, Ipswich MA, United States of America.
PLoS One. 2017 Sep 25;12(9):e0185340. doi: 10.1371/journal.pone.0185340. eCollection 2017.
Zika virus (ZIKV) has emerged as a major global public health concern in the last two years due to its link as a causative agent of human birth defects. Its rapid expansion into the Western Hemisphere as well as the ability to be transmitted from mother to fetus, through sexual transmission and possibly through blood transfusions has increased the need for a rapid and expansive public health response to this unprecedented epidemic. A non-invasive and rapid ZIKV diagnostic screening assay that can be performed in a clinical setting throughout pregnancy is vital for prenatal care of women living in areas of the world where exposure to the virus is possible. To meet this need we have developed a sensitive and specific reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay to detect ZIKV RNA in urine and serum with a simple visual detection. RT-LAMP results were shown to have a limit of detection 10-fold higher than qRT-PCR. As little as 1.2 RNA copies/μl was detected by RT-LAMP from a panel of 178 diagnostic specimens. The assay was shown to be highly specific for ZIKV RNA when tested with diagnostic specimens positive for dengue virus (DENV) and chikungunya virus (CHIKV). The assay described here illustrates the potential for a fast, reliable, sensitive and specific assay for the detection of ZIKV from urine or serum that can be performed in a clinical or field setting with minimal equipment and technological expertise.
寨卡病毒（ZIKV）在过去两年中已成为全球主要的公共卫生问题，因为它被认为是导致人类出生缺陷的病原体。它迅速蔓延至西半球，并且能够通过母婴传播、性传播以及可能通过输血传播，这使得针对这一前所未有的疫情迅速展开广泛的公共卫生应对措施变得更加必要。一种可在整个孕期临床环境中进行的非侵入性快速寨卡病毒诊断筛查检测方法，对于生活在可能接触该病毒地区的女性的产前护理至关重要。为满足这一需求，我们开发了一种灵敏且特异的逆转录环介导等温扩增（RT-LAMP）检测方法，通过简单的视觉检测来检测尿液和血清中的寨卡病毒RNA。结果显示，RT-LAMP的检测限比定量逆转录聚合酶链反应（qRT-PCR）高10倍。从178份诊断标本中，RT-LAMP检测到低至1.2个RNA拷贝/微升。当用登革热病毒（DENV）和基孔肯雅病毒（CHIKV）阳性的诊断标本进行测试时，该检测方法对寨卡病毒RNA显示出高度特异性。本文所述的检测方法表明，有可能开发出一种快速、可靠、灵敏且特异的检测方法，用于从尿液或血清中检测寨卡病毒，该方法可在临床或现场环境中使用最少的设备和技术专业知识进行操作。
