{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:14:25Z","timestamp":1790342065345,"version":"4.1.0"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Virol J"],"published-print":{"date-parts":[[2012,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Yellow Fever virus (YFV) is an important arboviral pathogen in much of sub-Saharan Africa and the tropical Americas. It is the prototype member of the genus <jats:italic>Flavivirus<\/jats:italic> and is transmitted primarily by <jats:italic>Aedes (Stegomyia)<\/jats:italic> mosquitoes. The incidence of human infections in endemic areas has risen in recent years. Prompt and dependable identification of YFV is a critical component of response to suspect cases.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>We developed a one-step SYBR Green I-based real-time quantitative RT-PCR (qRT-PCR) assay targeting the 5'NTR and capsid-gene junction--for rapid detection and quantification of YFV. The detection limit was 1 PFU\/mL, 10-fold more sensitive than conventional RT-PCR, and there was no cross-reactivity with closely related flaviviruses or with alphaviruses. Viral load in samples was determined by standard curve plotted from cycle threshold (Ct) values and virus concentration. The efficacy of the assay in mosquitoes was assessed with spiked samples. The utility of the assay for screening of pooled mosquitoes was also confirmed. Replication of a Cameroon isolate of YFV in <jats:italic>Ae. aegypti<\/jats:italic> revealed a marked variation in susceptibility among different colonies at different days post infection (pi).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>The SYBR Green-1 based qRT-PCR assay is a faster, simpler, more sensitive and less expensive procedure for detection and quantification of YFV than other currently used methods.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1743-422x-9-27","type":"journal-article","created":{"date-parts":[[2012,1,24]],"date-time":"2012-01-24T07:16:53Z","timestamp":1327389413000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Development of a SYBR green I based RT-PCR assay for yellow fever virus: application in assessment of YFV infection in Aedes aegypti"],"prefix":"10.1186","volume":"9","author":[{"given":"Paban Kumar","family":"Dash","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Boutonnier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric","family":"Prina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shashi","family":"Sharma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Reiter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2012,1,22]]},"reference":[{"key":"1700_CR1","volume-title":"Yellow fever fact sheet No 100","author":"WHO (World Health Organiszation)","year":"2009","unstructured":"WHO (World Health Organiszation): Yellow fever fact sheet No 100.2009. [http:\/\/www.who.int\/mediacentre\/factsheets\/fs100\/en\/]"},{"key":"1700_CR2","first-page":"591","volume-title":"MMWR","author":"JE Staples","year":"2010","unstructured":"Staples JE, Gershman M, Fischer M, Centers for Disease Control and Prevention: Yellow fever vaccine: recommendations of the advisory committee on immunization practices (ACIP). MMWR 2010, 591-27."},{"key":"1700_CR3","doi-asserted-by":"publisher","first-page":"1143","DOI":"10.1086\/516111","volume":"25","author":"JM McFarland","year":"1997","unstructured":"McFarland JM, Baddour LM, Nelson JE, Elkins SK, et al.: Imported yellow fever in a United States citizen. Clin Infect Dis 1997, 25: 1143-1147. 10.1086\/516111","journal-title":"Clin Infect Dis"},{"key":"1700_CR4","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1016\/j.jcv.2004.12.001","volume":"33","author":"HG Bae","year":"2005","unstructured":"Bae HG, Drosten C, Emmerich P, Colebunders R, et al.: Analysis of two imported cases of yellow fever infection from ivory coast and the Gambia to Germany and Belgium. J Clin Virol 2005, 33: 274-280. 10.1016\/j.jcv.2004.12.001","journal-title":"J Clin Virol"},{"key":"1700_CR5","doi-asserted-by":"publisher","first-page":"649","DOI":"10.1146\/annurev.mi.44.100190.003245","volume":"44","author":"TJ Chambers","year":"1990","unstructured":"Chambers TJ, Hahn CS, Galler R, Rice CM: Flavivirus genome organization, expression, and replication. Annu Rev Microbiol 1990, 44: 649-688. 10.1146\/annurev.mi.44.100190.003245","journal-title":"Annu Rev Microbiol"},{"key":"1700_CR6","doi-asserted-by":"publisher","first-page":"343","DOI":"10.1016\/S0065-3527(03)60009-6","volume":"60","author":"TP Monath","year":"2003","unstructured":"Monath TP, Barrett ADT: Pathogenesis and pathophysiology of yellow fever. Adv Virus Res 2003, 60: 343-395.","journal-title":"Adv Virus Res"},{"key":"1700_CR7","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1016\/j.jviromet.2010.05.006","volume":"168","author":"CF Yang","year":"2010","unstructured":"Yang CF, Chen CF, Su CL, Teng HJ, et al.: Screening of mosquitoes using SYBR green I-based real-time RT-PCR with group-specific primers for detection of flaviviruses and alphaviruses in Taiwan. J Virol Methods 2010, 168: 147-151. 10.1016\/j.jviromet.2010.05.006","journal-title":"J Virol Methods"},{"key":"1700_CR8","doi-asserted-by":"publisher","first-page":"1292","DOI":"10.1093\/nar\/30.6.1292","volume":"30","author":"IM Mackay","year":"2002","unstructured":"Mackay IM, Arden KE: Nitsche A. Real-time PCR in virology. Nucleic Acids Res 2002, 30: 1292-1305. 10.1093\/nar\/30.6.1292","journal-title":"Nucleic Acids Res"},{"key":"1700_CR9","doi-asserted-by":"publisher","first-page":"2323","DOI":"10.1128\/JCM.40.7.2323-2330.2002","volume":"40","author":"C Drosten","year":"2002","unstructured":"Drosten C, G\u00f6ttig S, Schilling S, Asper M, et al.: Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean-Congo hemorrhagic fever virus, rift valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR. J Clin Microbiol 2002, 40: 2323-2330. 10.1128\/JCM.40.7.2323-2330.2002","journal-title":"J Clin Microbiol"},{"key":"1700_CR10","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1016\/S0166-0934(03)00129-0","volume":"110","author":"HG Bae","year":"2003","unstructured":"Bae HG, Nitsche A, Teichmann A, Biel SS, et al.: Detection of yellow fever virus: a comparison of quantitative real-time PCR and plaque assay. J Virol Methods 2003, 110: 185-191. 10.1016\/S0166-0934(03)00129-0","journal-title":"J Virol Methods"},{"key":"1700_CR11","doi-asserted-by":"publisher","first-page":"584","DOI":"10.1128\/JCM.00842-06","volume":"45","author":"DY Chao","year":"2007","unstructured":"Chao DY, Davis BS, Chang GJ: Development of multiplex real-time reverse transcriptase PCR assays for detecting eight medically important flaviviruses in mosquitoes. J Clin Microbiol 2007, 45: 584-589. 10.1128\/JCM.00842-06","journal-title":"J Clin Microbiol"},{"key":"1700_CR12","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/j.jviromet.2008.03.026","volume":"151","author":"N Mantel","year":"2008","unstructured":"Mantel N, Aguirre M, Gulia S, Girerd-Chambaz Y, et al.: Standardized quantitative RT-PCR assays for quantitation of yellow fever and chimeric yellow fever-dengue vaccines. J Virol Methods 2008, 151: 40-46. 10.1016\/j.jviromet.2008.03.026","journal-title":"J Virol Methods"},{"key":"1700_CR13","doi-asserted-by":"publisher","first-page":"1511","DOI":"10.1128\/JCM.42.4.1511-1518.2004","volume":"42","author":"JF Papin","year":"2004","unstructured":"Papin JF, Vahrson W, Dittmer DP: SYBR green-based real-time quantitative PCR assay for detection of West Nile Virus circumvents false-negative results due to strain variability. J Clin Microbiol 2004, 42: 1511-1518. 10.1128\/JCM.42.4.1511-1518.2004","journal-title":"J Clin Microbiol"},{"key":"1700_CR14","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1586\/14737159.1.2.153","volume":"1","author":"M Schutten","year":"2001","unstructured":"Schutten M, Niesters HG: Clinical utility of viral quantification as a tool for disease monitoring. Expert Rev Mol Diagn 2001, 1: 153-162. 10.1586\/14737159.1.2.153","journal-title":"Expert Rev Mol Diagn"},{"key":"1700_CR15","doi-asserted-by":"publisher","first-page":"589","DOI":"10.1089\/vbz.2007.0266","volume":"8","author":"T Balenghien","year":"2008","unstructured":"Balenghien T, Vazeille M, Grandadam M, Schaffner F, et al.: Vector competence of some French Culex and Aedes mosquitoes for west Nile virus. Vector Borne Zoonotic Dis 2008, 8: 589-595. 10.1089\/vbz.2007.0266","journal-title":"Vector Borne Zoonotic Dis"},{"issue":"11","key":"1700_CR16","doi-asserted-by":"publisher","first-page":"e1168","DOI":"10.1371\/journal.pone.0001168","volume":"2","author":"M Vazeille","year":"2007","unstructured":"Vazeille M, Moutailler S, Coudrier D, Rousseaux C, et al.: Two Chikungunya isolates from the outbreak of La reunion (Indian ocean) exhibit different patterns of infection in the mosquito, Aedes albopictus. PLoS One 2007,2(11):e1168. 10.1371\/journal.pone.0001168","journal-title":"PLoS One"},{"key":"1700_CR17","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/j.jcv.2005.05.002","volume":"35","author":"DM Whiley","year":"2006","unstructured":"Whiley DM, Sloots TP: Sequence variation can affect the performance of minor groove binder TaqMan probes in viral diagnostic assays. J Clin Virol 2006, 35: 81-83. 10.1016\/j.jcv.2005.05.002","journal-title":"J Clin Virol"},{"key":"1700_CR18","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1016\/j.jcv.2010.04.013","volume":"48","author":"M Weidmann","year":"2010","unstructured":"Weidmann M, Faye O, Faye O, Kranaster R, et al.: Improved LNA probe-based assay for the detection of African and South American yellow fever virus strains. J Clin Virol 2010, 48: 187-192. 10.1016\/j.jcv.2010.04.013","journal-title":"J Clin Virol"},{"key":"1700_CR19","doi-asserted-by":"crossref","first-page":"132","DOI":"10.4269\/ajtmh.2006.74.132","volume":"74","author":"J Richardson","year":"2006","unstructured":"Richardson J, Molina-cruz A, Salazar MI, Black IV, et al.: Quantitative analysis of dengue-2 virus RNA during the extrinsic incubation period in individual Aedes aegypti. Am J Trop Med Hyg 2006, 74: 132-141.","journal-title":"Am J Trop Med Hyg"},{"key":"1700_CR20","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/1472-6785-10-8","volume":"10","author":"E Martin","year":"2010","unstructured":"Martin E, Moutailler S, Madec Y, Failloux A: Differential responses of the mosquito Aedes albopictus from the Indian ocean region to two Chikungunya isolates. BMC Ecology 2010, 10: 8. 10.1186\/1472-6785-10-8","journal-title":"BMC Ecology"},{"key":"1700_CR21","first-page":"173","volume":"71","author":"R Vicens","year":"1993","unstructured":"Vicens R, Robert V, Pignon D, Zeller H, et al.: Yellow fever epidemic in the extreme North of Cameroon in 1990: first yellow fever virus isolation in Cameroon. Bull World Health Organ 1993, 71: 173-176.","journal-title":"Bull World Health Organ"},{"key":"1700_CR22","first-page":"43","volume-title":"Virology: A Practical Approach","author":"EA Gould","year":"1991","unstructured":"Gould EA, Clegg TCS: Growth, assay and purification of Alphaviruses and Flaviviruses. In Virology: A Practical Approach. Edited by: Mahy BWJ. Oxford University Press; 1991:43-78."},{"key":"1700_CR23","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1128\/jcm.30.3.545-551.1992","volume":"30","author":"RS Lanciotti","year":"1992","unstructured":"Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, et al.: Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 1992, 30: 545-551.","journal-title":"J Clin Microbiol"},{"key":"1700_CR24","doi-asserted-by":"publisher","first-page":"1427","DOI":"10.3201\/eid1209.060200","volume":"12","author":"MM Parida","year":"2006","unstructured":"Parida MM, Dash PK, Tripathi NK, et al.: Japanese Encephalitis Outbreak, India, 2005. Emerg Infect Dis 2006, 12: 1427-1430. 10.3201\/eid1209.060200","journal-title":"Emerg Infect Dis"},{"key":"1700_CR25","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1089\/vbz.2006.0648","volume":"7","author":"PK Dash","year":"2007","unstructured":"Dash PK, Parida MM, Santhosh SR, Verma SK, et al.: East central south African genotype as the causative agent in reemergence of Chikungunya outbreak in India. Vector Borne Zoonotic Dis 2007, 7: 519-527. 10.1089\/vbz.2006.0648","journal-title":"Vector Borne Zoonotic Dis"},{"key":"1700_CR26","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1186\/1743-422X-5-20","volume":"5","author":"P Saxena","year":"2008","unstructured":"Saxena P, Dash PK, Santhosh SR, Shrivastava A, et al.: Development and evaluation of one step single tube multiplex RT-PCR for rapid detection and typing of dengue viruses. Virol J 2008, 5: 20. 10.1186\/1743-422X-5-20","journal-title":"Virol J"}],"container-title":["Virology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1743-422X-9-27.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T18:31:36Z","timestamp":1630521096000},"score":1,"resource":{"primary":{"URL":"https:\/\/virologyj.biomedcentral.com\/articles\/10.1186\/1743-422X-9-27"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,1,22]]},"references-count":26,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2012,12]]}},"alternative-id":["1700"],"URL":"https:\/\/doi.org\/10.1186\/1743-422x-9-27","relation":{},"ISSN":["1743-422X"],"issn-type":[{"value":"1743-422X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,1,22]]},"assertion":[{"value":"21 June 2011","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 January 2012","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 January 2012","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"27"}}