Detection of Encephalitis Viruses in Mosquitoes (Diptera: Culicidae) and Avian Tissues
Diagnostic assays for the detection of St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) viruses in mosquito pools and avian tissues were compared for sensitivity, accuracy and specificity. The in situ enzyme immunoassay (EIA), plaque assay on Vero cells, passage in Aedes albop...
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| Veröffentlicht in: | Journal of medical entomology 2002-03, Vol.39 (2), p.312-323 |
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| container_title | Journal of medical entomology |
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| creator | Kramer, L. D. Wolfe, T. M. Green, E. N. Chiles, R. E. Fallah, H. Fang, Y. Reisen, W. K. |
| description | Diagnostic assays for the detection of St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) viruses in mosquito pools and avian tissues were compared for sensitivity, accuracy and specificity. The in situ enzyme immunoassay (EIA), plaque assay on Vero cells, passage in Aedes albopictus Skuse C6/36 and C7/10 cells, antigen capture enzyme immunoassay (AC-EIA), and single and multiplex reverse transcription-polymerase chain reactions (RT-PCR) were evaluated using pools of 50 mosquitoes containing 1–2 experimentally infected individuals. RT-PCR was the most sensitive assay, with a detection limit of |
| doi_str_mv | 10.1603/0022-2585-39.2.312 |
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D. ; Wolfe, T. M. ; Green, E. N. ; Chiles, R. E. ; Fallah, H. ; Fang, Y. ; Reisen, W. K.</creator><creatorcontrib>Kramer, L. D. ; Wolfe, T. M. ; Green, E. N. ; Chiles, R. E. ; Fallah, H. ; Fang, Y. ; Reisen, W. K.</creatorcontrib><description>Diagnostic assays for the detection of St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) viruses in mosquito pools and avian tissues were compared for sensitivity, accuracy and specificity. The in situ enzyme immunoassay (EIA), plaque assay on Vero cells, passage in Aedes albopictus Skuse C6/36 and C7/10 cells, antigen capture enzyme immunoassay (AC-EIA), and single and multiplex reverse transcription-polymerase chain reactions (RT-PCR) were evaluated using pools of 50 mosquitoes containing 1–2 experimentally infected individuals. RT-PCR was the most sensitive assay, with a detection limit of <0.1 plaque forming unit. AC-EIA was the fastest and most economical procedure, but was the least sensitive, detecting only 38% of positive pools. The in situ EIA included initial virus amplification on Vero cells, thereby improving assay sensitivity to detect 68% of positive pools. Passage in C6/36 and/or C7/10 cell culture revealed the presence of infectious virus in samples positive by RT-PCR, but initially negative by plaque assay on Vero cell culture, indicating that detection was related to assay sensitivity and not to the absence of intact infectious virus. Combining WEE and SLE RT-PCR assays into a multiplex assay reduced sensitivity, but still detected viral RNA at titers below plaque assay sensitivity. Plaque assay on Vero cells, mosquito cell passage, and several RT-PCR procedures were evaluated for their ability to detect WEE and SLE in white-crowned sparrow tissues during acute and chronic stages of infection. All assays detected virus during acute infection at times of high viremia; however, only RT-PCR assays were positive by day 7 when virus was not detected in sera. RT-PCR detected SLE RNA in spleen tissue from one bird 51 d after infection. Assay sensitivity also was compared using extracts of homogenized bird organs spiked with known titers of WEE and SLE. Trizol RNA extraction followed by Qiagen one-step RT-PCR was the most sensitive method, but occasionally resulted in the presence of secondary bands confounding interpretation and requiring confirmatory assays. A balanced surveillance program should combine systems that allow the detection of new agents and the sensitive monitoring of endemic agents to provide an early warning of pending health risks.</description><identifier>ISSN: 0022-2585</identifier><identifier>EISSN: 1938-2928</identifier><identifier>DOI: 10.1603/0022-2585-39.2.312</identifier><identifier>PMID: 11931031</identifier><identifier>CODEN: JMENA6</identifier><language>eng</language><publisher>Lanham, MD: Entomological Society of America</publisher><subject>Aedes - virology ; Animals ; arboviruses ; Asian tiger mosquito ; Biological and medical sciences ; Bird Diseases - pathology ; Bird Diseases - virology ; Birds ; Cercopithecus aethiops ; Culex - virology ; DNA, Viral - analysis ; Encephalitis ; Encephalitis Virus, St. Louis - genetics ; Encephalitis Virus, St. Louis - immunology ; Encephalitis Virus, St. Louis - isolation & purification ; Encephalitis Virus, Western Equine - genetics ; Encephalitis Virus, Western Equine - immunology ; Encephalitis Virus, Western Equine - isolation & purification ; Encephalitis, St. Louis - pathology ; Encephalitis, St. Louis - veterinary ; Encephalitis, St. Louis - virology ; Encephalomyelitis, Western Equine - pathology ; Encephalomyelitis, Western Equine - veterinary ; Encephalomyelitis, Western Equine - virology ; enzyme immunoassay ; Enzymes ; Female ; Fundamental and applied biological sciences. Psychology ; Health aspects ; Medically important nuisances and vectors, pests of stored products and materials: population survey and control ; Mosquitoes ; reverse transcription-polymerase chain reaction ; RNA ; Sensitivity and Specificity ; Songbirds - virology ; surveillance ; Vectors. Intermediate hosts ; Vero Cells ; virus isolation</subject><ispartof>Journal of medical entomology, 2002-03, Vol.39 (2), p.312-323</ispartof><rights>Entomological Society of America</rights><rights>2002 INIST-CNRS</rights><rights>COPYRIGHT 2002 Oxford University Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b519t-3fb911de7c620b716382288a1ee545f4446cbb89ea75ebd8a5e10b4aa3db3ffe3</citedby><cites>FETCH-LOGICAL-b519t-3fb911de7c620b716382288a1ee545f4446cbb89ea75ebd8a5e10b4aa3db3ffe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://bioone.org/doi/pdf/10.1603/0022-2585-39.2.312$$EPDF$$P50$$Gbioone$$H</linktopdf><link.rule.ids>314,780,784,26978,27924,27925,52363</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=13561618$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11931031$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kramer, L. D.</creatorcontrib><creatorcontrib>Wolfe, T. M.</creatorcontrib><creatorcontrib>Green, E. N.</creatorcontrib><creatorcontrib>Chiles, R. E.</creatorcontrib><creatorcontrib>Fallah, H.</creatorcontrib><creatorcontrib>Fang, Y.</creatorcontrib><creatorcontrib>Reisen, W. K.</creatorcontrib><title>Detection of Encephalitis Viruses in Mosquitoes (Diptera: Culicidae) and Avian Tissues</title><title>Journal of medical entomology</title><addtitle>J Med Entomol</addtitle><description>Diagnostic assays for the detection of St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) viruses in mosquito pools and avian tissues were compared for sensitivity, accuracy and specificity. The in situ enzyme immunoassay (EIA), plaque assay on Vero cells, passage in Aedes albopictus Skuse C6/36 and C7/10 cells, antigen capture enzyme immunoassay (AC-EIA), and single and multiplex reverse transcription-polymerase chain reactions (RT-PCR) were evaluated using pools of 50 mosquitoes containing 1–2 experimentally infected individuals. RT-PCR was the most sensitive assay, with a detection limit of <0.1 plaque forming unit. AC-EIA was the fastest and most economical procedure, but was the least sensitive, detecting only 38% of positive pools. The in situ EIA included initial virus amplification on Vero cells, thereby improving assay sensitivity to detect 68% of positive pools. Passage in C6/36 and/or C7/10 cell culture revealed the presence of infectious virus in samples positive by RT-PCR, but initially negative by plaque assay on Vero cell culture, indicating that detection was related to assay sensitivity and not to the absence of intact infectious virus. Combining WEE and SLE RT-PCR assays into a multiplex assay reduced sensitivity, but still detected viral RNA at titers below plaque assay sensitivity. Plaque assay on Vero cells, mosquito cell passage, and several RT-PCR procedures were evaluated for their ability to detect WEE and SLE in white-crowned sparrow tissues during acute and chronic stages of infection. All assays detected virus during acute infection at times of high viremia; however, only RT-PCR assays were positive by day 7 when virus was not detected in sera. RT-PCR detected SLE RNA in spleen tissue from one bird 51 d after infection. Assay sensitivity also was compared using extracts of homogenized bird organs spiked with known titers of WEE and SLE. Trizol RNA extraction followed by Qiagen one-step RT-PCR was the most sensitive method, but occasionally resulted in the presence of secondary bands confounding interpretation and requiring confirmatory assays. A balanced surveillance program should combine systems that allow the detection of new agents and the sensitive monitoring of endemic agents to provide an early warning of pending health risks.</description><subject>Aedes - virology</subject><subject>Animals</subject><subject>arboviruses</subject><subject>Asian tiger mosquito</subject><subject>Biological and medical sciences</subject><subject>Bird Diseases - pathology</subject><subject>Bird Diseases - virology</subject><subject>Birds</subject><subject>Cercopithecus aethiops</subject><subject>Culex - virology</subject><subject>DNA, Viral - analysis</subject><subject>Encephalitis</subject><subject>Encephalitis Virus, St. Louis - genetics</subject><subject>Encephalitis Virus, St. Louis - immunology</subject><subject>Encephalitis Virus, St. Louis - isolation & purification</subject><subject>Encephalitis Virus, Western Equine - genetics</subject><subject>Encephalitis Virus, Western Equine - immunology</subject><subject>Encephalitis Virus, Western Equine - isolation & purification</subject><subject>Encephalitis, St. Louis - pathology</subject><subject>Encephalitis, St. Louis - veterinary</subject><subject>Encephalitis, St. Louis - virology</subject><subject>Encephalomyelitis, Western Equine - pathology</subject><subject>Encephalomyelitis, Western Equine - veterinary</subject><subject>Encephalomyelitis, Western Equine - virology</subject><subject>enzyme immunoassay</subject><subject>Enzymes</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Health aspects</subject><subject>Medically important nuisances and vectors, pests of stored products and materials: population survey and control</subject><subject>Mosquitoes</subject><subject>reverse transcription-polymerase chain reaction</subject><subject>RNA</subject><subject>Sensitivity and Specificity</subject><subject>Songbirds - virology</subject><subject>surveillance</subject><subject>Vectors. Intermediate hosts</subject><subject>Vero Cells</subject><subject>virus isolation</subject><issn>0022-2585</issn><issn>1938-2928</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkEFvFCEYhomxsWv1D3gwXDR6mJUPhhnwttlWbdKml9orAeZDMbPDFmZM_Pey2Y29Gg6EL8_L9-Yh5A2wNXRMfGKM84ZLJRuh13wtgD8jK9BCNVxz9Zys_gHn5GUpvxhjClr9gpxDpYAJWJGHS5zRzzFNNAV6NXnc_7RjnGOhDzEvBQuNE71N5XGJc6qvD5dxP2O2n-l2GaOPg8WP1E4D3fyOdqL3sZQFyytyFuxY8PXpviDfv1zdb781N3dfr7ebm8ZJ0HMjgtMAA_a-48z10AnFuVIWEGUrQ9u2nXdOabS9RDcoKxGYa60VgxMhoLgg74__7nN6rHtns4vF4zjaCdNSTA-y63SvKrg-gj_siCZOIc3Z-noG3EWfJgyxzjdSSJAcNK8Bfgz4nErJGMw-x53Nfwwwc9BvDnbNwa4R2nBT9dfQ21Odxe1weIqcfFfg3QmwxdsxZDv5WJ44ITvo4FCXHTkXUy33P7v_AmkjmwE</recordid><startdate>20020301</startdate><enddate>20020301</enddate><creator>Kramer, L. D.</creator><creator>Wolfe, T. M.</creator><creator>Green, E. N.</creator><creator>Chiles, R. E.</creator><creator>Fallah, H.</creator><creator>Fang, Y.</creator><creator>Reisen, W. K.</creator><general>Entomological Society of America</general><general>Oxford University Press</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20020301</creationdate><title>Detection of Encephalitis Viruses in Mosquitoes (Diptera: Culicidae) and Avian Tissues</title><author>Kramer, L. D. ; Wolfe, T. M. ; Green, E. N. ; Chiles, R. E. ; Fallah, H. ; Fang, Y. ; Reisen, W. 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Psychology</topic><topic>Health aspects</topic><topic>Medically important nuisances and vectors, pests of stored products and materials: population survey and control</topic><topic>Mosquitoes</topic><topic>reverse transcription-polymerase chain reaction</topic><topic>RNA</topic><topic>Sensitivity and Specificity</topic><topic>Songbirds - virology</topic><topic>surveillance</topic><topic>Vectors. Intermediate hosts</topic><topic>Vero Cells</topic><topic>virus isolation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kramer, L. D.</creatorcontrib><creatorcontrib>Wolfe, T. M.</creatorcontrib><creatorcontrib>Green, E. N.</creatorcontrib><creatorcontrib>Chiles, R. E.</creatorcontrib><creatorcontrib>Fallah, H.</creatorcontrib><creatorcontrib>Fang, Y.</creatorcontrib><creatorcontrib>Reisen, W. 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K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of Encephalitis Viruses in Mosquitoes (Diptera: Culicidae) and Avian Tissues</atitle><jtitle>Journal of medical entomology</jtitle><addtitle>J Med Entomol</addtitle><date>2002-03-01</date><risdate>2002</risdate><volume>39</volume><issue>2</issue><spage>312</spage><epage>323</epage><pages>312-323</pages><issn>0022-2585</issn><eissn>1938-2928</eissn><coden>JMENA6</coden><abstract>Diagnostic assays for the detection of St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) viruses in mosquito pools and avian tissues were compared for sensitivity, accuracy and specificity. The in situ enzyme immunoassay (EIA), plaque assay on Vero cells, passage in Aedes albopictus Skuse C6/36 and C7/10 cells, antigen capture enzyme immunoassay (AC-EIA), and single and multiplex reverse transcription-polymerase chain reactions (RT-PCR) were evaluated using pools of 50 mosquitoes containing 1–2 experimentally infected individuals. RT-PCR was the most sensitive assay, with a detection limit of <0.1 plaque forming unit. AC-EIA was the fastest and most economical procedure, but was the least sensitive, detecting only 38% of positive pools. The in situ EIA included initial virus amplification on Vero cells, thereby improving assay sensitivity to detect 68% of positive pools. Passage in C6/36 and/or C7/10 cell culture revealed the presence of infectious virus in samples positive by RT-PCR, but initially negative by plaque assay on Vero cell culture, indicating that detection was related to assay sensitivity and not to the absence of intact infectious virus. Combining WEE and SLE RT-PCR assays into a multiplex assay reduced sensitivity, but still detected viral RNA at titers below plaque assay sensitivity. Plaque assay on Vero cells, mosquito cell passage, and several RT-PCR procedures were evaluated for their ability to detect WEE and SLE in white-crowned sparrow tissues during acute and chronic stages of infection. All assays detected virus during acute infection at times of high viremia; however, only RT-PCR assays were positive by day 7 when virus was not detected in sera. RT-PCR detected SLE RNA in spleen tissue from one bird 51 d after infection. Assay sensitivity also was compared using extracts of homogenized bird organs spiked with known titers of WEE and SLE. Trizol RNA extraction followed by Qiagen one-step RT-PCR was the most sensitive method, but occasionally resulted in the presence of secondary bands confounding interpretation and requiring confirmatory assays. A balanced surveillance program should combine systems that allow the detection of new agents and the sensitive monitoring of endemic agents to provide an early warning of pending health risks.</abstract><cop>Lanham, MD</cop><pub>Entomological Society of America</pub><pmid>11931031</pmid><doi>10.1603/0022-2585-39.2.312</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Aedes - virology Animals arboviruses Asian tiger mosquito Biological and medical sciences Bird Diseases - pathology Bird Diseases - virology Birds Cercopithecus aethiops Culex - virology DNA, Viral - analysis Encephalitis Encephalitis Virus, St. Louis - genetics Encephalitis Virus, St. Louis - immunology Encephalitis Virus, St. Louis - isolation & purification Encephalitis Virus, Western Equine - genetics Encephalitis Virus, Western Equine - immunology Encephalitis Virus, Western Equine - isolation & purification Encephalitis, St. Louis - pathology Encephalitis, St. Louis - veterinary Encephalitis, St. Louis - virology Encephalomyelitis, Western Equine - pathology Encephalomyelitis, Western Equine - veterinary Encephalomyelitis, Western Equine - virology enzyme immunoassay Enzymes Female Fundamental and applied biological sciences. Psychology Health aspects Medically important nuisances and vectors, pests of stored products and materials: population survey and control Mosquitoes reverse transcription-polymerase chain reaction RNA Sensitivity and Specificity Songbirds - virology surveillance Vectors. Intermediate hosts Vero Cells virus isolation |
| title | Detection of Encephalitis Viruses in Mosquitoes (Diptera: Culicidae) and Avian Tissues |
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