{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,18]],"date-time":"2026-05-18T00:32:22Z","timestamp":1779064342780,"version":"3.51.4"},"reference-count":27,"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":[[2008,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Hepatitis A virus (HAV), the causative agent of acute hepatitis in humans, is an atypical Picornaviridae that grows poorly in cell culture. HAV titrations are laborious and time-consuming because the virus in general does not cause cytopathic effect and is detected by immunochemical or molecular probes. Simple HAV titration assays could be developed using currently available viral construct containing selectable markers.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>We developed an antibiotic resistance titration assay (ARTA) based on the infection of human hepatoma cells with a wild type HAV construct containing a blasticidin (Bsd) resistance gene. Human hepatoma cells infected with the HAV-Bsd construct survived selection with 2 \u03bcg\/ml of blasticidin whereas uninfected cells died within a few days. At 8 days postinfection, the color of the pH indicator phenol red in cell culture media correlated with the presence of HAV-Bsd-infected blasticidin-resistant cells: an orange-to-yellow color indicated the presence of growing cells whereas a pink-to-purple color indicated that the cells were dead. HAV-Bsd titers were determined by an endpoint dilution assay based on the color of the cell culture medium scoring orange-to-yellow wells as positive and pink-to-purple wells as negative for HAV. As a proof-of-concept, we used the ARTA to evaluate the HAV neutralization potency of two commercially available human immune globulin (IG) preparations and a WHO International Standard for anti-HAV. The three IG preparations contained comparable levels of anti-HAV antibodies that neutralized approximately 1.5 log of HAV-Bsd. Similar neutralization results were obtained in the absence of blasticidin by an endpoint dilution ELISA at 2 weeks postinfection.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>The ARTA is a simple and rapid method to determine HAV titers without using HAV-specific probes. We determined the HAV neutralization potency of human IG preparations in 8 days by ARTA compared to the 14 days required by the endpoint dilution ELISA. The ARTA reduced the labour, time, and cost of HAV titrations making it suitable for high throughput screening of sera and antivirals, determination of anti-HAV antibodies in human immune globulin preparations, and research applications that involve the routine evaluation of HAV titers.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1743-422x-5-155","type":"journal-article","created":{"date-parts":[[2008,12,18]],"date-time":"2008-12-18T19:16:09Z","timestamp":1229627769000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["A simple and rapid Hepatitis A Virus (HAV) titration assay based on antibiotic resistance of infected cells: evaluation of the HAV neutralization potency of human immune globulin preparations"],"prefix":"10.1186","volume":"5","author":[{"given":"Krishnamurthy","family":"Konduru","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Luisa","family":"Virata-Theimer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mei-ying W","family":"Yu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerardo G","family":"Kaplan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2008,12,18]]},"reference":[{"key":"504_CR1","unstructured":"Hepatitis A vaccines. World Health Organization Weekly Epidemiological Record 2000, 75: 38-44."},{"key":"504_CR2","doi-asserted-by":"crossref","first-page":"145","DOI":"10.4315\/0362-028X-70.1.145","volume":"70","author":"YC Shieh","year":"2007","unstructured":"Shieh YC, Khudyakov YE, Xia G, Ganova-Raeva LM, Khambaty FM, Woods JW, Veazey JE, Motes ML, Glatzer MB, Bialek SR, Fiore AE: Molecular confirmation of oysters as the vector for hepatitis A in a 2005 multistate outbreak. J Food Prot 2007, 70: 145-150.","journal-title":"J Food Prot"},{"key":"504_CR3","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1056\/NEJM199902253400802","volume":"340","author":"YJ Hutin","year":"1999","unstructured":"Hutin YJ, Pool V, Cramer EH, Nainan OV, Weth J, Williams IT, Goldstein ST, Gensheimer KF, Bell BP, Shapiro CN, et al.: A multistate, foodborne outbreak of hepatitis A. National Hepatitis A Investigation Team. N Engl J Med 1999, 340: 595-602. 10.1056\/NEJM199902253400802","journal-title":"N Engl J Med"},{"key":"504_CR4","doi-asserted-by":"publisher","first-page":"518","DOI":"10.1093\/infdis\/166.3.518","volume":"166","author":"MT Niu","year":"1992","unstructured":"Niu MT, Polish LB, Robertson BH, Khanna BK, Woodruff BA, Shapiro CN, Miller MA, Smith JD, Gedrose JK, Alter MJ, et al.: Multistate outbreak of hepatitis A associated with frozen strawberries. J Infect Dis 1992, 166: 518-524.","journal-title":"J Infect Dis"},{"key":"504_CR5","doi-asserted-by":"publisher","first-page":"890","DOI":"10.1056\/NEJMoa050855","volume":"353","author":"C Wheeler","year":"2005","unstructured":"Wheeler C, Vogt TM, Armstrong GL, Vaughan G, Weltman A, Nainan OV, Dato V, Xia G, Waller K, Amon J, et al.: An outbreak of hepatitis A associated with green onions. N Engl J Med 2005, 353: 890-897. 10.1056\/NEJMoa050855","journal-title":"N Engl J Med"},{"key":"504_CR6","doi-asserted-by":"publisher","first-page":"490","DOI":"10.1086\/499816","volume":"42","author":"M Mutsch","year":"2006","unstructured":"Mutsch M, Spicher VM, Gut C, Steffen R: Hepatitis A virus infections in travelers, 1988\u20132004. Clin Infect Dis 2006, 42: 490-497. 10.1086\/499816","journal-title":"Clin Infect Dis"},{"key":"504_CR7","unstructured":"Hepatitis Surveillance Report No. 61. 2005 Atlanta, GA: U.S. Department of Health and Human Services. Centers for Disease Control and Prevention; 2006."},{"key":"504_CR8","unstructured":"Prevention of hepatitis A through active or passive immunization: recommendations of the Advisory Committee on Immunization Practices (ACIP) Morbid Mortal Wkly Rep (MMWR) 2007, 1080-1084."},{"key":"504_CR9","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/oxfordjournals.aje.a120845","volume":"87","author":"JW Mosley","year":"1968","unstructured":"Mosley JW, Reisler DM, Brachott D, Roth D, Weiser J: Comparison of two lots of immune serum globulin for prophylaxis of infectious hepatitis. Am J Epidemiol 1968, 87: 539-550.","journal-title":"Am J Epidemiol"},{"key":"504_CR10","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1056\/NEJM199208133270702","volume":"327","author":"A Werzberger","year":"1992","unstructured":"Werzberger A, Mensch B, Kuter B, Brown L, Lewis J, Sitrin R, Miller W, Shouval D, Wiens B, Calandra G, et al.: A controlled trial of a formalin-inactivated hepatitis A vaccine in healthy children. N Engl J Med 1992, 327: 453-457.","journal-title":"N Engl J Med"},{"key":"504_CR11","doi-asserted-by":"publisher","first-page":"1685","DOI":"10.1056\/NEJMoa070546","volume":"357","author":"JC Victor","year":"2007","unstructured":"Victor JC, Monto AS, Surdina TY, Suleimenova SZ, Vaughan G, Nainan OV, Favorov MO, Margolis HS, Bell BP: Hepatitis A vaccine versus immune globulin for postexposure prophylaxis. N Engl J Med 2007, 357: 1685-1694. 10.1056\/NEJMoa070546","journal-title":"N Engl J Med"},{"key":"504_CR12","doi-asserted-by":"publisher","first-page":"580","DOI":"10.1093\/clinids\/14.2.580","volume":"14","author":"PL Winokur","year":"1992","unstructured":"Winokur PL, Stapleton JT: Immunoglobulin prophylaxis for hepatitis A. Clin Infect Dis 1992, 14: 580-586.","journal-title":"Clin Infect Dis"},{"key":"504_CR13","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1007\/978-1-59259-702-4_2","volume-title":"Viral Hepatitis: Diagnosis, Therapy and Prevention","author":"JT Stapleton","year":"1999","unstructured":"Stapleton JT: Hepatitis A: biology, pathogenesis, epidemiology, clinical description, and diagnosis. In Viral Hepatitis: Diagnosis, Therapy and Prevention. Edited by: Spector S. Humana Press, Totowa, NJ; 1999:7-33."},{"key":"504_CR14","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1002\/jmv.1890220107","volume":"22","author":"T Cromeans","year":"1987","unstructured":"Cromeans T, Sobsey MD, Fields HA: Development of a plaque assay for a cytopathic, rapidly replicating isolate of hepatitis A virus. J Med Virol 1987, 22: 45-56. 10.1002\/jmv.1890220107","journal-title":"J Med Virol"},{"key":"504_CR15","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1002\/jmv.1890220106","volume":"22","author":"DA Anderson","year":"1987","unstructured":"Anderson DA: Cytopathology, plaque assay, and heat inactivation of hepatitis A virus strain HM175. J Med Virol 1987, 22: 35-44. 10.1002\/jmv.1890220106","journal-title":"J Med Virol"},{"key":"504_CR16","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1128\/jcm.17.5.834-839.1983","volume":"17","author":"SM Lemon","year":"1983","unstructured":"Lemon SM, Binn LN, Marchwicki RH: Radioimmunofocus assay for quantitation of hepatitis A virus in cell cultures. J Clin Microbiol 1983, 17: 834-839.","journal-title":"J Clin Microbiol"},{"key":"504_CR17","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.jviromet.2005.10.023","volume":"133","author":"NA Counihan","year":"2005","unstructured":"Counihan NA, Daniel LM, Chojnacki J, Anderson DA: Infrared fluorescent immunofocus assay (IR-FIFA) for the quantitation of non-cytopathic and minimally cytopathic viruses. J Virol Methods 2005, 133: 62-69. 10.1016\/j.jviromet.2005.10.023","journal-title":"J Virol Methods"},{"key":"504_CR18","doi-asserted-by":"publisher","first-page":"1033","DOI":"10.1093\/infdis\/148.6.1033","volume":"148","author":"SM Lemon","year":"1983","unstructured":"Lemon SM, Binn LN: Serum neutralizing antibody response to hepatitis A virus. J Infect Dis 1983, 148: 1033-1039.","journal-title":"J Infect Dis"},{"key":"504_CR19","doi-asserted-by":"publisher","first-page":"2497","DOI":"10.1073\/pnas.84.8.2497","volume":"84","author":"JI Cohen","year":"1987","unstructured":"Cohen JI, Rosenblum B, Ticehurst JR, Daemer RJ, Feinstone SM, Purcell RH: Complete nucleotide sequence of an attenuated hepatitis A virus: comparison with wild-type virus. Proc Natl Acad Sci USA 1987, 84: 2497-2501. 10.1073\/pnas.84.8.2497","journal-title":"Proc Natl Acad Sci USA"},{"key":"504_CR20","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1128\/jvi.68.1.148-157.1994","volume":"68","author":"AW Funkhouser","year":"1994","unstructured":"Funkhouser AW, Purcell RH, D'Hondt E, Emerson SU: Attenuated hepatitis A virus: genetic determinants of adaptation to growth in MRC-5 cells. J Virol 1994, 68: 148-157.","journal-title":"J Virol"},{"key":"504_CR21","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1128\/JVI.75.2.717-725.2001","volume":"75","author":"E Silberstein","year":"2001","unstructured":"Silberstein E, Dveksler G, Kaplan GG: Neutralization of hepatitis A virus (HAV) by an immunoadhesin containing the cysteine-rich region of HAV cellular receptor-1. J Virol 2001, 75: 717-725. 10.1128\/JVI.75.2.717-725.2001","journal-title":"J Virol"},{"key":"504_CR22","doi-asserted-by":"publisher","first-page":"8765","DOI":"10.1128\/JVI.77.16.8765-8774.2003","volume":"77","author":"E Silberstein","year":"2003","unstructured":"Silberstein E, Xing L, Beek W, Lu J, Cheng H, Kaplan GG: Alteration of hepatitis A virus (HAV) particles by a soluble form of HAV cellular receptor 1 containing the immunoglobin-and mucin-like regions. J Virol 2003, 77: 8765-8774.","journal-title":"J Virol"},{"key":"504_CR23","doi-asserted-by":"publisher","first-page":"2950","DOI":"10.1128\/JVI.79.5.2950-2955.2005","volume":"79","author":"DA Feigelstock","year":"2005","unstructured":"Feigelstock DA, Thompson P, Kaplan GG: Growth of hepatitis A virus in a mouse liver cell line. J Virol 2005, 79: 2950-2955. 10.1128\/JVI.79.5.2950-2955.2005","journal-title":"J Virol"},{"key":"504_CR24","doi-asserted-by":"publisher","first-page":"1352","DOI":"10.1128\/JVI.80.3.1352-1360.2006","volume":"80","author":"K Konduru","year":"2006","unstructured":"Konduru K, Kaplan GG: Stable growth of wild-type hepatitis A virus in cell culture. J Virol 2006, 80: 1352-1360. 10.1128\/JVI.80.3.1352-1360.2006","journal-title":"J Virol"},{"key":"504_CR25","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1002\/jmv.10457","volume":"71","author":"J Graff","year":"2003","unstructured":"Graff J, Emerson SU: Importance of amino acid 216 in nonstructural protein 2B for replication of hepatitis A virus in cell culture and in vivo. J Med Virol 2003, 71: 7-17. 10.1002\/jmv.10457","journal-title":"J Med Virol"},{"key":"504_CR26","first-page":"493","volume":"27","author":"LJ Reed","year":"1938","unstructured":"Reed LJ, Muench H: A simple method of estimating fifty per cent end points. Am J Hyg 1938, 27: 493-497.","journal-title":"Am J Hyg"},{"key":"504_CR27","first-page":"411","volume":"54","author":"RJ Gerety","year":"1983","unstructured":"Gerety RJ, Smallwood LA, Finlayson JS, Tabor E: Standardization of the antibody to hepatitis A virus (anti-HAV) content of immunoglobulin. Dev Biol Stand 1983, 54: 411-416.","journal-title":"Dev Biol Stand"}],"container-title":["Virology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1743-422X-5-155.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T09:51:08Z","timestamp":1630489868000},"score":1,"resource":{"primary":{"URL":"https:\/\/virologyj.biomedcentral.com\/articles\/10.1186\/1743-422X-5-155"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,12]]},"references-count":27,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2008,12]]}},"alternative-id":["504"],"URL":"https:\/\/doi.org\/10.1186\/1743-422x-5-155","relation":{},"ISSN":["1743-422X"],"issn-type":[{"value":"1743-422X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,12]]},"assertion":[{"value":"5 December 2008","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 December 2008","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 December 2008","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"155"}}