O'Guinn Monica L, Lee John S, Kondig John P, Fernandez Roberto, Carbajal Faustino
United States Army Center For Health Promotion and Preventative Medicine-Pacific, Camp Zama, Japan.
Am J Trop Med Hyg. 2004 Feb;70(2):164-71.
In support of efforts to develop rapid diagnostic assays for use in the field, reverse transcription-polymerase chain reaction (RT-PCR) assays were developed to detect arboviruses circulating in the Amazon Basin region of Peru. Previous knowledge of arthropod/pathogen relationships allowed a focused evaluation to be conducted in November 2000 that assessed the feasibility and reliability of a mobile, rapid, field-expedient RT-PCR diagnostic system aimed at detecting eastern equine encephalitis virus (EEEV) in Culex (Melanoconion) pedroi mosquitoes. Modifications were made to a commercially available mobile molecular laboratory kit and assay procedures were tailored for use under harsh environmental conditions with field-collected and field-processed mosquitoes. From CO2 baited mosquito light traps, 3,227 Cx. (Mel.) pedroi mosquitoes were collected and sorted into 117 pools. The pools were processed and assayed in the field by RT-PCR and five of those pools were found positive for EEEV. Laboratory sequence analysis confirmed the presence of two distinct subtypes of EEEV.
为支持开发用于现场的快速诊断检测方法，人们开发了逆转录聚合酶链反应（RT-PCR）检测方法，以检测在秘鲁亚马逊河流域地区传播的虫媒病毒。基于此前对节肢动物/病原体关系的了解，2000年11月进行了一次重点评估，评估一种移动、快速、便于现场操作的RT-PCR诊断系统检测库蚊（黑蚊亚属）中东部马脑炎病毒（EEEV）的可行性和可靠性。对市售的移动分子实验室试剂盒进行了改进，并针对在恶劣环境条件下对野外采集和野外处理的蚊子进行检测，对检测程序进行了调整。通过二氧化碳诱蚊灯诱捕器，共收集到3227只库蚊（黑蚊亚属），并分成117组。这些组在野外通过RT-PCR进行处理和检测，其中5组被发现EEEV呈阳性。实验室序列分析证实存在两种不同的EEEV亚型。
