Detection of Wuchereria bancrofti L3 larvae in mosquitoes: a reverse transcriptase PCR assay evaluating infection and infectivity.
Detection of Wuchereria bancrofti L3 larvae in mosquitoes: a reverse transcriptase PCR assay evaluating infection and infectivity.
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DOI：
10.1371/journal.pntd.0000602
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发表时间：
2010-02-16
影响因子：
3.8
通讯作者：
Williams SA
中科院分区：
文献类型：
--
作者：
Laney SJ;Ramzy RM;Helmy HH;Farid HA;Ashour AA;Weil GJ;Williams SA
用PCR检测蚊子体内的丝虫DNA不能区分感染性蚊子和受感染的蚊子。为了评估传播风险，需要一种可以特异性检测感染性L3期寄生虫的测定法。我们现在报告的发展，专门检测班氏吴策线虫在蚊子的感染阶段的测定。该检测试剂盒通过逆转录酶PCR（RT-PCR）检测L3激活的mRNA转录物。 W.使用生物信息学选择班氏表皮相关基因，并筛选其作为蚊子中L3检测的潜在诊断靶基因。使用RT-PCR对从喂食受感染血液后两周内每天收集的蚊子中分离的RNA进行表达谱测定。传统的多重RT-PCR和实时多重RT-PCR检测开发使用L3激活角质蛋白转录L3检测和组成型表达的转录本，tph-1，为“任何阶段”的检测。该方法可同时检测W。在合并的病媒蚊子中的班氏感染期幼虫和“任意期”幼虫。这种测试可能是有用的工具，用于评估在消除丝虫病计划的背景下，传播潜力的变化。淋巴丝虫病是一种由蚊子传播的寄生虫引起的致残和毁容疾病。寄生虫的生命周期需要两个宿主：蚊子载体和人类宿主。寄生虫的发育生命周期的一部分发生在蚊子体内，另一部分发生在人类宿主体内。这种寄生虫在蚊子体内经过四个阶段发展，只有最后一个阶段对人类具有感染性。第三幼虫阶段（L3）是感染阶段，当感染性蚊子叮咬人类时，会引发人类感染。目前有一个全球计划，试图通过向受影响的社区施用药物来消除这种疾病，目的是阻断寄生虫的传播。本文描述的新诊断工具使用分子技术来特异性地检测蚊子中寄生虫的感染阶段。可以一次检测许多蚊子，以评估社区中丝虫病持续传播的风险。此外，这种新的L3检测方法可以同时检测蚊子是否含有“任何阶段”的寄生虫。这提供了社区中人类感染率的信息。这两条信息都可用于评估消除疾病工作的进展情况。
Detection of filarial DNA in mosquitoes by PCR cannot differentiate infective mosquitoes from infected mosquitoes. In order to evaluate transmission risk an assay is needed that can specifically detect infective L3 stage parasites. We now report the development of an assay that specifically detects the infective stage of Wuchereria bancrofti in mosquitoes. The assay detects an L3-activated mRNA transcript by reverse-transcriptase PCR (RT-PCR). W. bancrofti cuticle-related genes were selected using bioinformatics and screened as potential diagnostic target genes for L3 detection in mosquitoes. Expression profiles were determined using RT-PCR on RNA isolated from mosquitoes collected daily across a two-week period after feeding on infected blood. Conventional multiplex RT-PCR and real-time multiplex RT-PCR assays were developed using an L3-activated cuticlin transcript for L3 detection and a constitutively expressed transcript, tph-1, for ‘any-stage’ detection. This assay can be used to simultaneously detect W. bancrofti infective stage larvae and ‘any-stage’ larvae in pooled vector mosquitoes. This test may be useful as a tool for assessing changes in transmission potential in the context of filariasis elimination programs. Lymphatic filariasis is a disabling and disfiguring disease caused by a parasite that is transmitted by a mosquito. The life cycle of the parasite requires two hosts: the mosquito vector and the human host. Part of the developmental life cycle of the parasite occurs in the mosquito and the other part in the human host. The parasite develops through four stages in the mosquito, only the last of which is infectious to humans. The third larval stage (L3) is the infective stage that initiates human infections when infective mosquitoes bite humans. There is currently a global program attempting to eliminate this disease by administering drugs to affected communities with the goal of interrupting transmission of the parasite. The new diagnostic tool described in this paper uses molecular techniques to specifically detect the infective stage of the parasite in mosquitoes. Many mosquitoes can be tested at one time to assess the risk of ongoing transmission of filariasis in communities. In addition, this new L3-detection assay can simultaneously detect whether the mosquitoes contain ‘any-stage’ of the parasite. This provides information on infection rates in humans in the community. Both pieces of information can be used in assessing the progress of disease elimination efforts.
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PCR和解剖作为监测法氏多尼西斯蚊子的丝状感染的工具。
DOI：
10.1186/1475-2883-5-2
发表时间：
2006-02-24
期刊：
Filaria journal
影响因子：
--
作者：
Plichart, Catherine;Sechan, Yves;Davies, Neil;Legrand, Anne-Marie
通讯作者：
Legrand, Anne-Marie
影响因子：
3.8
作者：
Ottesen, Eric A.;Hooper, Pamela J.;Biswas, Gautam
通讯作者：
Biswas, Gautam
DOI：
10.4269/ajtmh.2007.76.502
发表时间：
2007-03-01
影响因子：
3.3
作者：
Fischer, Peter;Erickson, Sara M.;Weil, Gary J.
通讯作者：
Weil, Gary J.
影响因子：
168.9
作者：
Ramzy, RMR;El Setouhy, M;Weil, GJ
通讯作者：
Weil, GJ
DOI：
10.1016/s0169-4758(97)01151-4
发表时间：
1997-12-01
期刊：
PARASITOLOGY TODAY
影响因子：
--
作者：
Michael, E;Bundy, DAP
通讯作者：
Bundy, DAP