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应用聚合酶链反应-酶联免疫吸附测定法检测巴布亚新几内亚丝虫病防治地区野生捕获的斑点按蚊群体中的班氏吴策线虫。

Application of a polymerase chain reaction-ELISA to detect Wuchereria bancrofti in pools of wild-caught Anopheles punctulatus in a filariasis control area in Papua New Guinea.

作者信息

Bockarie M J, Fischer P, Williams S A, Zimmerman P A, Griffin L, Alpers M P, Kazura J W

机构信息

Papua New Guinea Institute of Medical Research, Madang.

出版信息

Am J Trop Med Hyg. 2000 Mar;62(3):363-7. doi: 10.4269/ajtmh.2000.62.363.

DOI:10.4269/ajtmh.2000.62.363
PMID:11037778
Abstract

Chemotherapy-based eradication programs are aimed at stopping transmission of Wuchereria bancrofti by its obligatory mosquito vector. This study compares one year post-treatment W. bancrofti infection rates of Anopheles punctulatus, the main vector of lymphatic filariasis in Papua New Guinea, using traditional dissection techniques and a polymerase chain reaction (PCR)-based ELISA of a parasite-specific Ssp I repeat. A total of 633 mosquitoes in 35 batches were dissected. Six batches contained W. bancrofti-infected mosquitoes, giving a minimum infection rate of 0.9%. This value was not different than the actual infection rate, which was 9 (1.4%) of 633 mosquitoes (P = 0.48). The DNA was extracted from 47 pools containing a mean of 13.2 mosquitoes per pool. A total of 621 mosquitoes were processed for the PCR-ELISA, including 486 caught by human bait and 135 by light trap, which included both dead and live mosquitoes. Of 23 pools of alcohol-preserved human-bait mosquitoes, seven were positive by the PCR-ELISA, giving an infection rate identical to that obtained by dissection of individual mosquitoes (1.4%). The minimum infection rates for pools of light-trap mosquitoes found dead and alive were 2.7% (2 of 74) and 4.9% (3 of 61), respectively. These values did not differ from each other (P = 0.84), but the overall infection rate of light-trap mosquitoes was greater than that of mosquitoes captured by human bait (3.7% versus 1.4%; P = 0.09). These data indicate that the PCR-ELISA of a W. bancrofti Ssp I repeat using pools of mosquitoes is comparable to traditional dissection techniques for monitoring transmission intensity following introduction of mass chemotherapy. This approach may also be useful for rapid and cost-effective assessment of transmission in endemic areas where the frequency of overt lymphatic pathology is low.

摘要

基于化疗的根除计划旨在通过其必需的蚊媒阻止班氏吴策线虫的传播。本研究使用传统解剖技术和基于聚合酶链反应(PCR)的寄生虫特异性Ssp I重复序列酶联免疫吸附测定(ELISA),比较了巴布亚新几内亚淋巴丝虫病主要传播媒介微小按蚊在治疗后一年的班氏吴策线虫感染率。共解剖了35批633只蚊子。6批含有感染班氏吴策线虫的蚊子,最低感染率为0.9%。该值与实际感染率无差异,实际感染率为633只蚊子中的9只(1.4%)(P = 0.48)。从47个样本池中提取DNA,每个样本池平均含有13.2只蚊子。共对621只蚊子进行了PCR-ELISA检测,其中包括486只人饵诱捕的蚊子和135只光诱捕的蚊子,光诱捕的蚊子包括死蚊和活蚊。在23个用酒精保存的人饵诱捕蚊子样本池中,7个通过PCR-ELISA呈阳性,感染率与解剖单个蚊子获得的感染率相同(1.4%)。光诱捕的死蚊和活蚊样本池的最低感染率分别为2.7%(74只中的2只)和4.9%(61只中的3只)。这些值彼此无差异(P = 0.84),但光诱捕蚊子的总体感染率高于人饵诱捕的蚊子(3.7%对1.4%;P = 0.09)。这些数据表明,使用蚊子样本池对班氏吴策线虫Ssp I重复序列进行PCR-ELISA检测,在引入大规模化疗后监测传播强度方面与传统解剖技术相当。这种方法对于明显淋巴病理学频率较低的流行地区的传播快速且经济高效评估也可能有用。

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