Detection of human herpesvirus-7 by qualitative nested-PCR: comparison between healthy individuals and liver transplant recipients

Abstract

Diagnosis of human herpesvirus-7 active infection in transplant patients has proved difficult, because this virus is ubiquitous and can cause persistent infections in the host. The significance of viral DNA detected in leukocytes by PCR is unclear and cross-reaction in serological tests may occur. This study aimed to evaluate nested-PCR to detect human herpesvirus-7 active infection in liver transplant recipients compared to healthy individuals. human herpesvirus-7 nested-PCR was performed on leukocytes and sera of 53 healthy volunteers and sera of 29 liver transplant recipients. In healthy volunteers, human herpesvirus-7 was detected in 28.3% of leukocytes and 0% of serum. human herpesvirus-7 was detected in sera of 48.2% of the liver transplant recipients. Nested-PCR on DNA extracted from leukocytes detected latent infection and the study suggests that nested-PCR performed on serum could be useful to detect human herpesvirus-7 active infection in liver transplant recipients.Key-words: human herpesvirus-7. healthy individuals. Nested-polymerase chain reaction. Liver transplantation. RESUMODiagnóstico da infecção ativa pelo herpesvirus humano-7 é difícil devido ao fato deste vírus ser ubíquo e poder causar infecção persistente no hospedeiro. O significado da detecção do DNA viral por reação em cadeia da polimerase não é claro e, reações cruzadas podem ocorrer em testes sorológicos. O objetivo deste estudo foi avaliar a nested-PCR para detectar infecção ativa pelo herpesvirus-7 em receptores hepáticos comparando com indivíduos sadios. Nested-PCR para herpesvirus-7 foi realizado em leucócitos e soro de 53 voluntários sadios e em soro de 29 receptores hepáticos. Nos voluntários sadios, herpesvirus-7 foi detectado em 28,3% de leucócitos e 0% de soro. herpesvirus-7 foi detectado em soro de 48,2% de receptores hepáticos. Nested-PCR em DNA extraído de leucócitos detectou infecção latente e o estudo sugere que nested-PCR realizada em soro poderia ser útil para detectar infecção ativa por herpesvirus-7 em receptores de fígado.Palavras-chaves: herpesvirus humano-7. Indivíduos sadios. Nested-reação em cadeia da polimerase. Transplante hepático. human herpesvirus-7 (hhV-7) was first isolated by Frenkel et al 5 from activated CD4 + peripheral blood T cells of a healthy individual. It is a member of the betaherpesvirinae subfamily of the Betaherpesviridae (DNA virus). Both, hhV-7 and hhV-6 (human herpesvirus 6) primary infections cause common febrile infectious syndromes of early childhood, known as exanthem subitum and roseola 16 . Investigations conducted in the United States 8 and Mexico 11 presented hhV-7 seroprevalence rates of 65% and 98%, respectively. In Brazil, Freitas et al 4 found a hhV-7 seroprevalence rate of 93.3% in individuals > 10 years of age.Similar to other betaherpesviruses, hhV-7 frequently remains latent in the host and can reactivate during immunosuppression following organ transplantation 9 . The most well-known member of the betaherpesviruses is human cytomegalovirus (hCMV) and it is consider...

Active human herpesvirus infections in adults with systemic lupus erythematosus and correlation with the SLEDAI score

“…HHV coinfection was considered in case of two or more positive HHV-DNA results in the same serum sample. The following isolated and/or combined symptoms were used to characterize active HHV infection, and/or likely HCMV disease: fever, headache, abdominal pain, seizure, changes in behavior, low levels of consciousness, mental confusion, motor dysfunction, neuropathy, paresthesia, drowsiness, vomiting, weakness, weight loss, myoclonus, memory loss, visual and psychomotor impairment, genital and orofacial herpesvirus, ocular infection, dermatitis, varicella, mononucleosis syndrome, Burkitt's lymphoma, nasopharyngeal carcinoma, Hodgkin's disease, HCMV colitis, retinitis, hepatitis, pneumonitis, roseola infantum or exanthem subitum, Kaposi's sarcoma, primary effusion lymphoma and Cattleman's disease [27][28][29][30][31][32].…”

Human herpesvirus‐7 in Brazilian liver transplant recipients: a follow‐up comparison between molecular and immunological assays

“…However, nested PCR carried out in DNA extracted from serum can be considered more speci¢c than PCR carried out in DNA extract from whole blood or peripheral blood leukocytes (9). A previous study conducted in our center showed that nested PCR in serum, according to the method described in this paper, did not detect latent infection in a healthy cohort (12). Serum samples that presented b-globin gene ampli¢cation were excluded from this study, and the presence of HHV-7 DNA in serum as a result of cell lysis is therefore unlikely.…”

Betaherpesviruses in Adult Liver Transplant Recipients

“…In addition, IgM against HHV-7 was found in all samples contributing to the hypothesis of true active infection had occurred. Since that this method did not detect latent infection in previously study (Thomasini et al, 2008) and blood was collected before surgery, it could be explained by reactivation caused by underlying liver disease or by transient viremia. Although some syndromes related to HHV-7 in immunocompetent patients have been described (Ward et al, 2005), studies in pre-transplant time should be performed to evaluate each hypothesis.…”

“…In our center, we found that nested-PCR carried out in DNA extracted from sera did not detect latent HHV-7 in a healthy cohort (Thomasini et al, 2008). In addition, positive IgM anti-HHV-7 and/or significant increase in IgG anti-HHV-7 titers were correlated with positive nested-PCR for HHV-7 in adult liver transplant recipients (Peigo et al, 2009).…”