Development of a novel dengue virus serotype-specific multiplex real-time reverse transcription-polymerase chain reaction assay for blood screening.
Development of a novel dengue virus serotype-specific multiplex real-time reverse transcription-polymerase chain reaction assay for blood screening.
复制标题
开发一种用于血液筛查的新型登革热病毒血清型特异性多重实时逆转录聚合酶链反应测定法。
DOI：
10.1111/trf.13875
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发表时间：
2016
期刊：
影响因子：
2.9
通讯作者：
and Isao Hamaguchi
中科院分区：
文献类型：
--
作者：
Kenta Tezuka;Madoka Kuramitsu;Kazu Okuma;Kiyoko Nojima;Kumiko Araki;Naoya Shinohara;Chieko Matsumoto;Masahiro Satake;Tomohiko Takasaki;Masayuki Saijo;Ichiro Kurane;and Isao Hamaguchi
登革热是由黄病毒属的四种相关RNA病毒引起的，登革热病毒（DENV）-1，-2，-3和-4，它们通过蚊子传播给人类。虽然DENV在日本不是地方性的，但2014年发生了本土登革热疫情。在DENV流行国家，在使用血液和血浆制品后也报告了几例输血传播病例。本研究的目的是开发一种新的多重逆转录-聚合酶链反应（RT-PCR）检测DENV血液screening.Study设计和方法进行大规模的寡核苷酸筛选，以获得DENV特异性引物和探针使用各种DENV临床分离株。利用所筛选的寡核苷酸建立了检测登革病毒RNA的多重RT-PCR方法，并对该方法检测登革病毒RNA的能力进行了评价，确定了适合检测登革病毒RNA的寡核苷酸的数量，成功建立了一种新的登革病毒血清型特异性多重RT-PCR方法。结论建立的DENV血清型特异性多重RT-PCR检测方法简便、灵敏、定量，可应用于血液标本的筛查，以预防输血传播的DENV感染。
BACKGROUNDDengue fever is caused by four related RNA viruses of the genusFlavivirus, dengue virus (DENV)‐1, ‐2, ‐3, and ‐4, which are transmitted to humans by mosquitoes. Although DENV is not endemic in Japan, an autochthonous dengue outbreak occurred in 2014. Several transfusion‐transmitted cases have also been reported after the use of blood and plasma products in DENV‐endemic countries. The aim of this study was to develop a novel multiplex reverse transcription–polymerase chain reaction (RT‐PCR) assay for DENV blood screening.STUDY DESIGN AND METHODSLarge‐scale oligonucleotide screening was performed to obtain DENV‐specific primers and probes using a variety of DENV clinical isolates. A multiplex RT‐PCR assay was then developed using the identified oligonucleotides and the ability of this assay to detect DENV RNA was evaluated.RESULTSA number of oligonucleotides suitable for DENV RNA detection were identified and a novel DENV serotype‐specific multiplex RT‐PCR assay was successfully established. Comparative analysis revealed that the multiplex assay could detect levels of viral contamination as low as 100 viral copies/mL.CONCLUSIONThis established serotype‐specific multiplex RT‐PCR assay provides a simple, sensitive, and quantitative detection method for DENV, which could be applied in the screening of blood samples to prevent transfusion‐transmitted DENV infection.