ANTIGEN-CAPTURE ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR DETECTION AND QUANTIFICATION OF CRIMEAN-CONGO HEMORRHAGIC-FEVER VIRUS IN THE TICK, HYALOMMA-TRUNCATUM
ANTIGEN-CAPTURE ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR DETECTION AND QUANTIFICATION OF CRIMEAN-CONGO HEMORRHAGIC-FEVER VIRUS IN THE TICK, HYALOMMA-TRUNCATUM
复制标题
DOI：
10.1016/0166-0934(93)90174-p
复制
发表时间：
1993-04-01
影响因子：
3.1
通讯作者：
KSIAZEK, TG
中科院分区：
文献类型：
--
作者：
LOGAN, TM;LINTHICUM, KJ;KSIAZEK, TG
将病毒抗原捕获ELISA与人细胞单层上的病毒空斑试验进行比较，以检测和定量研磨的实验感染的截形璃眼蜱中的克里米亚-刚果出血热病毒。在蜱幼虫暴露于病毒血症小鼠的悬浮液中，ELISA检测到13%阳性，而空斑试验检测到3%（n = 721）阳性。成年蜱接种病毒，并采样到102天后，84%的ELISA阳性相比，36%（n = 273）阳性的空斑试验。在病毒接种后两周内，两种试验检测到的阳性蜱比例相似;然而，在接种后18-102天，ELISA在100%的接种成年蜱中呈阳性，而空斑试验在33%（n = 135）的相同标本中呈阳性。通过ELISA检测，在10%（n = 101）的第一代子代中检测到CCHF病毒抗原（作为未喂养幼虫池），但空斑试验未检测到病毒，表明通过ELISA在这些蜱后代中检测到非感染性病毒片段或极低水平的活病毒。通过空斑试验检测，接种病毒的未进食成年蜱在接种后第8天几乎全部感染;在接种后第21天，仅33%被检测为阳性。从接种后第21-31天开始，允许这些蜱虫的队列吸血。在喂食后进行检测时，所有雌性蜱和近50%的雄性蜱均被检测为病毒阳性。这表明病毒可能在未进食蜱中持续存在，低于空斑试验的可检测水平，并在进食血液的蜱中增加至可检测水平。然而，从接种后第14天开始，在整个研究的剩余时间内，ELISA检测到100%的蜱虫为病毒阳性，无论饲养状态如何（接种后第102天）。提示抗原检测ELISA法检测蜱体内CCHF病毒的灵敏度高于空斑法。由于受感染的蜱虫可能在其余生中通过ELISA保持抗原阳性，因此该检测方法将是蜱虫对CCHF病毒的现场调查和媒介能力研究的重大改进。
A viral antigen-capture ELISA was compared to a viral plaque-assay on human cell monolayers for detection and quantification of Crimean-Congo hemorrhagic fever virus in triturated experimentally infected Hyalomma truncatum ticks. In suspensions of ticks exposed as larvae to viremic mice, the ELISA detected 13% positive as compared to 3% (n = 721) positive by plaque-assay. Adult ticks inoculated with virus and sampled up to 102 days later were 84% positive by ELISA compared to 36% (n = 273) positive by plaque-assay. The two tests detected similar proportions of positive ticks in the two weeks immediately after viral inoculation; however, the ELISA was positive in 100% of inoculated adult ticks from 18-102 days post-inoculation while the plaque-assay was positive in 33% (n = 135) of the same specimens. CCHF viral antigen was detected in 10% (n = 101) of first-generation progeny tested as unfed larva pools by ELISA, yet no virus was detected by plaque-assay, indicating that either non-infective viral fragments or very low levels of live virus were detected by ELISA in these tick progeny. As detected by plaque assay, virus inoculated unfed adult ticks were virtually all infected by day 8 post-inoculation; by day 21 post-inoculation only 33% were detected as positive. A cohort of these ticks were allowed to blood feed from day 21-31 post-inoculation. When assayed after feeding all female ticks and nearly 50% of male ticks were detected as virus-positive. This indicates that the virus likely persisted in the unfed ticks below the level of detectability of the plaque-assay and increased in the blood fed ticks up to a detectable level. The ELISA however, detected 100% of ticks as virus-positive from day 14 post-inoculation throughout the remainder of the study, regardless of feeding status (day 102 post-inoculation). These results indicate that antigen-detection ELISA is more sensitive in detecting CCHF virus in ticks than plaque-assay. Since an infected tick remains antigen-positive by ELISA for possibly the remainder of its life, this assay will be a major improvement in field surveys and vector competency studies of ticks for CCHF virus.