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Published in: BMC Infectious Diseases 1/2019

Open Access 01-12-2019 | Herpes Virus | Research article

Multiplex qPCR facilitates identification of betaherpesviruses in patients with acute liver failure of unknown etiology

Authors: Jéssica Vasques Raposo, Arthur Daniel Rocha Alves, Alexandre dos Santos da Silva, Damião Carlos dos Santos, Juliana Gil Melgaço, Otacílio C. Moreira, Marcelo Alves Pinto, Vanessa Salete de Paula

Published in: BMC Infectious Diseases | Issue 1/2019

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Abstract

Background

The etiology of acute liver failure (ALF) is often unknown and reported to be associated with herpesviruses in a number of cases. In this study, we examined for betaherpesviruses infections in patients with ALF of unknown etiology using a multiplex qPCR to Betaherpesviruses subfamily.

Methods

Liver explant and serum samples from 27 patients with ALF of unknown etiology were analyzed with the aid of multiplex qPCR to identify betaherpesviruses. All positive samples were sequenced to confirm herpes infection and liver enzyme levels evaluated.

Results

Betaherpesviruses infection was effectively detected using multiplex qPCR. Six (22%) HHV-6, one (3%) HCMV and two (7%) dual infections (one with HHV-7/HHV-6, and the other with HHV-7/ HCMV). Interestingly, HHV-7 was only detected in the presence of other betaherpesviruses. Sequencing information confirmed betaherpesviruses infection. High hepatic enzyme levels and INR values> 1.5 were determined in all betaherpesvirus-positive patients.

Conclusions

Multiplex qPCR facilitated efficient quantification, indicating that differentiation between betaherpesviruses is possible with the sole use of real-time PCR. Liver explant and serum samples were positive for some betaherpesviruses, and coinfection of HHV-7 with HHV-6 and HCMV was additionally detected. Based on these results, we propose that ALF patients should be screened for the presence of betaherpesviruses.
Literature
1.
go back to reference Mochida S, Takikawa Y, Nakayama N, Oketani M, Naiki T, Yamagishi Y, et al. Diagnostic criteria of acute liver failure: A report by the Intractable Hepato-Biliary Diseases Study Group of Japan. Hepatol Res. 2011;41(9):805–12.CrossRef Mochida S, Takikawa Y, Nakayama N, Oketani M, Naiki T, Yamagishi Y, et al. Diagnostic criteria of acute liver failure: A report by the Intractable Hepato-Biliary Diseases Study Group of Japan. Hepatol Res. 2011;41(9):805–12.CrossRef
2.
go back to reference Ehlers B, Borchers K, Grund C, Frolich K, Ludwig H, Buhk HJ. Detection of new DNA polymerase genes of known and potentially novel herpesviruses by PCR with degenerate and deoxyinosine-substituted primers. Virus Genes. 1999;18(3):211–20.CrossRef Ehlers B, Borchers K, Grund C, Frolich K, Ludwig H, Buhk HJ. Detection of new DNA polymerase genes of known and potentially novel herpesviruses by PCR with degenerate and deoxyinosine-substituted primers. Virus Genes. 1999;18(3):211–20.CrossRef
3.
go back to reference Abbo L, Alcaide ML, Pano JR, Robinson PG, Campo RE. Fulminant hepatitis from herpes simplex virus type 2 in an immunocompetent adult. Transpl Infect Dis. 2007;9(4):323–6.CrossRef Abbo L, Alcaide ML, Pano JR, Robinson PG, Campo RE. Fulminant hepatitis from herpes simplex virus type 2 in an immunocompetent adult. Transpl Infect Dis. 2007;9(4):323–6.CrossRef
4.
go back to reference Ishikawa K, Hasegawa K, Naritomi T, Kanai N, Ogawa M, Kato Y, et al. Prevalence of herpesviridae and hepatitis virus sequences in the livers of patients with fulminant hepatitis of unknown etiology in Japan. J Gastroenterol. 2002;37(7):523–30.CrossRef Ishikawa K, Hasegawa K, Naritomi T, Kanai N, Ogawa M, Kato Y, et al. Prevalence of herpesviridae and hepatitis virus sequences in the livers of patients with fulminant hepatitis of unknown etiology in Japan. J Gastroenterol. 2002;37(7):523–30.CrossRef
5.
go back to reference Lacoste V, Verschoor EJ, Nerrienet E, Gessain A. A novel homologue of human herpesvirus 6 in chimpanzees. J Gen Virol. 2005;86(Pt 8):2135–40.CrossRef Lacoste V, Verschoor EJ, Nerrienet E, Gessain A. A novel homologue of human herpesvirus 6 in chimpanzees. J Gen Virol. 2005;86(Pt 8):2135–40.CrossRef
6.
go back to reference Clark DA. Human herpesvirus 6 and human herpesvirus 7: emerging pathogens in transplant patients. Int J Hematol. 2002;76(Suppl 2):246–52.CrossRef Clark DA. Human herpesvirus 6 and human herpesvirus 7: emerging pathogens in transplant patients. Int J Hematol. 2002;76(Suppl 2):246–52.CrossRef
7.
go back to reference Cisneros-Herreros JM, Herrero-Romero M. Hepatitis due to herpes group viruses. Enferm Infecc Microbiol Clin. 2006;24(6):392–7 quiz 8.CrossRef Cisneros-Herreros JM, Herrero-Romero M. Hepatitis due to herpes group viruses. Enferm Infecc Microbiol Clin. 2006;24(6):392–7 quiz 8.CrossRef
8.
go back to reference Harma M, Hockerstedt K, Krogerus L, Lautenschlager I. Pretransplant human herpesvirus 6 infection of patients with acute liver failure is a risk factor for posttransplant human herpesvirus 6 infection of the liver. Transplantation. 2006;81(3):367–72.CrossRef Harma M, Hockerstedt K, Krogerus L, Lautenschlager I. Pretransplant human herpesvirus 6 infection of patients with acute liver failure is a risk factor for posttransplant human herpesvirus 6 infection of the liver. Transplantation. 2006;81(3):367–72.CrossRef
9.
go back to reference Paula VS, Silva AP, Michel D, Melgaco JG, Vieira YR, Santos DC, et al. Acute liver failure in an immunocompetent patient. J Clin Virol. 2014;60(1):1–3.CrossRef Paula VS, Silva AP, Michel D, Melgaco JG, Vieira YR, Santos DC, et al. Acute liver failure in an immunocompetent patient. J Clin Virol. 2014;60(1):1–3.CrossRef
10.
go back to reference Fernandez-Ruiz M, Kumar D, Husain S, Lilly L, Renner E, Mazzulli T, et al. Utility of a monitoring strategy for human herpesviruses 6 and 7 viremia after liver transplantation: a randomized clinical trial. Transplantation. 2015;99(1):106–13.CrossRef Fernandez-Ruiz M, Kumar D, Husain S, Lilly L, Renner E, Mazzulli T, et al. Utility of a monitoring strategy for human herpesviruses 6 and 7 viremia after liver transplantation: a randomized clinical trial. Transplantation. 2015;99(1):106–13.CrossRef
11.
go back to reference Yu YD, Park GC, Park PJ, Choi YI, Hwang S, Song GW, et al. Cytomegalovirus infection-associated fulminant hepatitis in an immunocompetent adult requiring emergency living-donor liver transplantation: report of a case. Surg Today. 2013;43(4):424–8.CrossRef Yu YD, Park GC, Park PJ, Choi YI, Hwang S, Song GW, et al. Cytomegalovirus infection-associated fulminant hepatitis in an immunocompetent adult requiring emergency living-donor liver transplantation: report of a case. Surg Today. 2013;43(4):424–8.CrossRef
12.
go back to reference Charnot-Katsikas A, Baewer D, Cook L, David MZ. Fulminant hepatic failure attributed to infection with human herpesvirus 6 (HHV-6) in an immunocompetent woman: A case report and review of the literature. J Clin Virol. 2016;75:27–32.CrossRef Charnot-Katsikas A, Baewer D, Cook L, David MZ. Fulminant hepatic failure attributed to infection with human herpesvirus 6 (HHV-6) in an immunocompetent woman: A case report and review of the literature. J Clin Virol. 2016;75:27–32.CrossRef
13.
go back to reference Arribas Anta J, Zaera de la Fuente C, Graus Morales J, Lopez Duran S, Canete Ruiz A, Gea Rodriguez F, et al. Fulminant hepatitis caused by herpes virus 6 in immunocompetent adults. Gastroenterol Hepatol. 2016;39(8):533–4.CrossRef Arribas Anta J, Zaera de la Fuente C, Graus Morales J, Lopez Duran S, Canete Ruiz A, Gea Rodriguez F, et al. Fulminant hepatitis caused by herpes virus 6 in immunocompetent adults. Gastroenterol Hepatol. 2016;39(8):533–4.CrossRef
14.
go back to reference Burd EM. Validation of laboratory-developed molecular assays for infectious diseases. Clin Microbiol Rev. 2010;23(3):550–76.CrossRef Burd EM. Validation of laboratory-developed molecular assays for infectious diseases. Clin Microbiol Rev. 2010;23(3):550–76.CrossRef
15.
go back to reference Fellner MD, Durand K, Rodriguez M, Irazu L, Alonio V, Picconi MA. Duplex realtime PCR method for Epstein-Barr virus and human DNA quantification: its application for post-transplant lymphoproliferative disorders detection. Braz J Infect Dis. 2014;18(3):271–80.CrossRef Fellner MD, Durand K, Rodriguez M, Irazu L, Alonio V, Picconi MA. Duplex realtime PCR method for Epstein-Barr virus and human DNA quantification: its application for post-transplant lymphoproliferative disorders detection. Braz J Infect Dis. 2014;18(3):271–80.CrossRef
16.
go back to reference Fernandes-Monteiro AG, Trindade GF, Yamamura AM, Moreira OC, de Paula VS, Duarte AC, et al. New approaches for the standardization and validation of a real-time qPCR assay using TaqMan probes for quantification of yellow fever virus on clinical samples with high quality parameters. Hum Vaccin Immunother. 2015;11(7):1865–71.CrossRef Fernandes-Monteiro AG, Trindade GF, Yamamura AM, Moreira OC, de Paula VS, Duarte AC, et al. New approaches for the standardization and validation of a real-time qPCR assay using TaqMan probes for quantification of yellow fever virus on clinical samples with high quality parameters. Hum Vaccin Immunother. 2015;11(7):1865–71.CrossRef
17.
go back to reference Riediger C, Sauer P, Matevossian E, Muller MW, Buchler P, Friess H. Herpes simplex virus sepsis and acute liver failure. Clin Transpl. 2009;23(Suppl 21):37–41.CrossRef Riediger C, Sauer P, Matevossian E, Muller MW, Buchler P, Friess H. Herpes simplex virus sepsis and acute liver failure. Clin Transpl. 2009;23(Suppl 21):37–41.CrossRef
18.
go back to reference Sassenscheidt J, Rohayem J, Illmer T, Bandt D. Detection of beta-herpesviruses in allogenic stem cell recipients by quantitative real-time PCR. J Virol Methods. 2006;138(1–2):40–8.CrossRef Sassenscheidt J, Rohayem J, Illmer T, Bandt D. Detection of beta-herpesviruses in allogenic stem cell recipients by quantitative real-time PCR. J Virol Methods. 2006;138(1–2):40–8.CrossRef
19.
go back to reference Hara S, Kimura H, Hoshino Y, Tanaka N, Nishikawa K, Ihira M, et al. Detection of herpesvirus DNA in the serum of immunocompetent children. Microbiol Immunol. 2002;46(3):177–80.CrossRef Hara S, Kimura H, Hoshino Y, Tanaka N, Nishikawa K, Ihira M, et al. Detection of herpesvirus DNA in the serum of immunocompetent children. Microbiol Immunol. 2002;46(3):177–80.CrossRef
20.
go back to reference Nishiwaki M, Fujimuro M, Teishikata Y, Inoue H, Sasajima H, Nakaso K, et al. Epidemiology of Epstein-Barr virus, cytomegalovirus, and Kaposi's sarcoma-associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay. J Med Virol. 2006;78(12):1635–42.CrossRef Nishiwaki M, Fujimuro M, Teishikata Y, Inoue H, Sasajima H, Nakaso K, et al. Epidemiology of Epstein-Barr virus, cytomegalovirus, and Kaposi's sarcoma-associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay. J Med Virol. 2006;78(12):1635–42.CrossRef
21.
go back to reference Al Midani A, Pinney J, Field N, Atkinson C, Haque T, Harber M. Fulminant hepatitis following primary herpes simplex virus infection. Saudi J Kidney Dis Transpl. 2011;22(1):107–11.PubMed Al Midani A, Pinney J, Field N, Atkinson C, Haque T, Harber M. Fulminant hepatitis following primary herpes simplex virus infection. Saudi J Kidney Dis Transpl. 2011;22(1):107–11.PubMed
22.
go back to reference Gallegos-Orozco JF, Rakela-Brodner J. Hepatitis viruses: not always what it seems to be. Rev Med Chil. 2010;138(10):1302–11.CrossRef Gallegos-Orozco JF, Rakela-Brodner J. Hepatitis viruses: not always what it seems to be. Rev Med Chil. 2010;138(10):1302–11.CrossRef
23.
go back to reference Herman D, Han H. Cytomegalovirus in liver transplant recipients. Curr Opin Organ Transplant. 2017;22(4):345–50.CrossRef Herman D, Han H. Cytomegalovirus in liver transplant recipients. Curr Opin Organ Transplant. 2017;22(4):345–50.CrossRef
24.
go back to reference Davalos F, Chaucer B, Zafar W, Salman S, Nfonoyim J. Dasatinib-induced CMV hepatitis in an immunocompetent patient: a rare complication of a common drug. Transl Oncol. 2016;9(3):248–50.CrossRef Davalos F, Chaucer B, Zafar W, Salman S, Nfonoyim J. Dasatinib-induced CMV hepatitis in an immunocompetent patient: a rare complication of a common drug. Transl Oncol. 2016;9(3):248–50.CrossRef
25.
go back to reference Mutimer DJ, Shaw J, O'Donnell K, Elias E. Enhanced (cytomegalovirus) viral replication after transplantation for fulminant hepatic failure. Liver Transpl Surg. 1997;3(5):506–12.CrossRef Mutimer DJ, Shaw J, O'Donnell K, Elias E. Enhanced (cytomegalovirus) viral replication after transplantation for fulminant hepatic failure. Liver Transpl Surg. 1997;3(5):506–12.CrossRef
26.
go back to reference Qian JY, Bai XY, Feng YL, Zhu WJ, Yao F, Li JN, et al. Cholestasis, ascites and pancytopenia in an immunocompetent adult with severe cytomegalovirus hepatitis. World J Gastroenterol. 2015;21(43):12505–9.CrossRef Qian JY, Bai XY, Feng YL, Zhu WJ, Yao F, Li JN, et al. Cholestasis, ascites and pancytopenia in an immunocompetent adult with severe cytomegalovirus hepatitis. World J Gastroenterol. 2015;21(43):12505–9.CrossRef
27.
go back to reference Asano Y, Yoshikawa T, Suga S, Yazaki T, Kondo K, Yamanishi K. Fatal fulminant hepatitis in an infant with human herpesvirus-6 infection. Lancet. 1990;335:862–3.CrossRef Asano Y, Yoshikawa T, Suga S, Yazaki T, Kondo K, Yamanishi K. Fatal fulminant hepatitis in an infant with human herpesvirus-6 infection. Lancet. 1990;335:862–3.CrossRef
28.
go back to reference Katsafanas GC, Schirmer EC, Wyatt LS, Frenkel N. In vitro activation of human herpesviruses 6 and 7 from latency. Proc Natl Acad Sci U S A. 1996;93(18):9788–92.CrossRef Katsafanas GC, Schirmer EC, Wyatt LS, Frenkel N. In vitro activation of human herpesviruses 6 and 7 from latency. Proc Natl Acad Sci U S A. 1996;93(18):9788–92.CrossRef
29.
go back to reference Lautenschlager I, Lappalainen M, Linnavuori K, Suni J, Hockerstedt K. CMV infection is usually associated with concurrent HHV-6 and HHV-7 antigenemia in liver transplant patients. J Clin Virol. 2002;25(Suppl 2):S57–61.CrossRef Lautenschlager I, Lappalainen M, Linnavuori K, Suni J, Hockerstedt K. CMV infection is usually associated with concurrent HHV-6 and HHV-7 antigenemia in liver transplant patients. J Clin Virol. 2002;25(Suppl 2):S57–61.CrossRef
30.
go back to reference Mendez JC, Dockrell DH, Espy MJ, Smith TF, Wilson JA, Harmsen WS, et al. Human beta-herpesvirus interactions in solid organ transplant recipients. J Infect Dis. 2001;183(2):179–84.CrossRef Mendez JC, Dockrell DH, Espy MJ, Smith TF, Wilson JA, Harmsen WS, et al. Human beta-herpesvirus interactions in solid organ transplant recipients. J Infect Dis. 2001;183(2):179–84.CrossRef
31.
go back to reference Pan H, Feng DN, Song L, Sun LR. Acute myeloid leukemia following etoposide therapy for EBV-associated hemophagocytic lymphohistiocytosis: a case report and a brief review of the literature. BMC Pediatr. 2016;16:116.CrossRef Pan H, Feng DN, Song L, Sun LR. Acute myeloid leukemia following etoposide therapy for EBV-associated hemophagocytic lymphohistiocytosis: a case report and a brief review of the literature. BMC Pediatr. 2016;16:116.CrossRef
32.
go back to reference Harma M, Hockerstedt K, Lyytikainen O, Lautenschlager I. HHV-6 and HHV-7 antigenemia related to CMV infection after liver transplantation. J Med Virol. 2006;78(6):800–5.CrossRef Harma M, Hockerstedt K, Lyytikainen O, Lautenschlager I. HHV-6 and HHV-7 antigenemia related to CMV infection after liver transplantation. J Med Virol. 2006;78(6):800–5.CrossRef
33.
go back to reference Kidd IM, Clark DA, Sabin CA, Andrew D, Hassan-Walker AF, Sweny P, et al. Prospective study of human betaherpesviruses after renal transplantation: association of human herpesvirus 7 and cytomegalovirus co-infection with cytomegalovirus disease and increased rejection. Transplantation. 2000;69(11):2400–4.CrossRef Kidd IM, Clark DA, Sabin CA, Andrew D, Hassan-Walker AF, Sweny P, et al. Prospective study of human betaherpesviruses after renal transplantation: association of human herpesvirus 7 and cytomegalovirus co-infection with cytomegalovirus disease and increased rejection. Transplantation. 2000;69(11):2400–4.CrossRef
34.
go back to reference Pellett Madan R, Hand J. Human herpesvirus 6, 7, and 8 in solid organ transplantation: Guidelines from the American Society of Transplantation Infectious Diseases Community of Practice. Clin Transplant. 2019:e13518. Pellett Madan R, Hand J. Human herpesvirus 6, 7, and 8 in solid organ transplantation: Guidelines from the American Society of Transplantation Infectious Diseases Community of Practice. Clin Transplant. 2019:e13518.
35.
go back to reference MOTTA VT. Bioquímica Clínica: Princípios e Interpretações. 5th ed; 2008. MOTTA VT. Bioquímica Clínica: Princípios e Interpretações. 5th ed; 2008.
36.
go back to reference Parker ZM, Pasieka TJ, Parker GA, Leib DA. Immune- and Nonimmune-Compartment-Specific Interferon Responses Are Critical Determinants of Herpes Simplex Virus-Induced Generalized Infections and Acute Liver Failure. J Virol. 2016;90(23):10789–99.CrossRef Parker ZM, Pasieka TJ, Parker GA, Leib DA. Immune- and Nonimmune-Compartment-Specific Interferon Responses Are Critical Determinants of Herpes Simplex Virus-Induced Generalized Infections and Acute Liver Failure. J Virol. 2016;90(23):10789–99.CrossRef
37.
go back to reference Somasekar S, Lee D, Rule J, Naccache SN, Stone M, Busch MP, et al. Viral surveillance in serum samples from patients with acute liver failure by metagenomic next-generation sequencing. Clin Infect Dis. 2017;65(9):1477–85.CrossRef Somasekar S, Lee D, Rule J, Naccache SN, Stone M, Busch MP, et al. Viral surveillance in serum samples from patients with acute liver failure by metagenomic next-generation sequencing. Clin Infect Dis. 2017;65(9):1477–85.CrossRef
38.
go back to reference Lee WM, Squires RH Jr, Nyberg SL, Doo E, Hoofnagle JH. Acute liver failure: summary of a workshop. Hepatology. 2008;47(4):1401–15.CrossRef Lee WM, Squires RH Jr, Nyberg SL, Doo E, Hoofnagle JH. Acute liver failure: summary of a workshop. Hepatology. 2008;47(4):1401–15.CrossRef
39.
go back to reference Squires RH Jr, Shneider BL, Bucuvalas J, Alonso E, Sokol RJ, Narkewicz MR, et al. Acute liver failure in children: the first 348 patients in the pediatric acute liver failure study group. J Pediatr. 2006;148(5):652–8.CrossRef Squires RH Jr, Shneider BL, Bucuvalas J, Alonso E, Sokol RJ, Narkewicz MR, et al. Acute liver failure in children: the first 348 patients in the pediatric acute liver failure study group. J Pediatr. 2006;148(5):652–8.CrossRef
40.
go back to reference Razonable RR. Human herpesviruses 6, 7 and 8 in solid organ transplant recipients. Am J Transplant. 2013;13(Suppl 3):67–77 quiz −8.CrossRef Razonable RR. Human herpesviruses 6, 7 and 8 in solid organ transplant recipients. Am J Transplant. 2013;13(Suppl 3):67–77 quiz −8.CrossRef
41.
go back to reference Ogata M. Human herpesvirus-6-associated diseases in hematopoietic stem cell transplantation: an update. Rinsho Ketsueki. 2016;57(3):298–306.PubMed Ogata M. Human herpesvirus-6-associated diseases in hematopoietic stem cell transplantation: an update. Rinsho Ketsueki. 2016;57(3):298–306.PubMed
42.
go back to reference Dandamudi R, Smith J, Dharnidharka VR. Renal transplantation and predisposition to opportunistic infections. Curr Opin Pediatr. 2019;31(2):226–31.CrossRef Dandamudi R, Smith J, Dharnidharka VR. Renal transplantation and predisposition to opportunistic infections. Curr Opin Pediatr. 2019;31(2):226–31.CrossRef
43.
go back to reference De Bolle L, Naesens L, De Clercq E. Update on human herpesvirus 6 biology, clinical features, and therapy. Clin Microbiol Rev. 2005;18(1):217–45.CrossRef De Bolle L, Naesens L, De Clercq E. Update on human herpesvirus 6 biology, clinical features, and therapy. Clin Microbiol Rev. 2005;18(1):217–45.CrossRef
Metadata
Title
Multiplex qPCR facilitates identification of betaherpesviruses in patients with acute liver failure of unknown etiology
Authors
Jéssica Vasques Raposo
Arthur Daniel Rocha Alves
Alexandre dos Santos da Silva
Damião Carlos dos Santos
Juliana Gil Melgaço
Otacílio C. Moreira
Marcelo Alves Pinto
Vanessa Salete de Paula
Publication date
01-12-2019
Publisher
BioMed Central
Published in
BMC Infectious Diseases / Issue 1/2019
Electronic ISSN: 1471-2334
DOI
https://doi.org/10.1186/s12879-019-4309-4

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