Multiplex qPCR Facilitates Identification of Betaherpesviruses in Patients With Acute Liver Failure of Unknown Etiology

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 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. Seven (26%), two (7%) and three (11%) cases were positive for HHV-6, HHV-7 and HCMV, respectively. Two cases of dual infection (HHV-7/HHV-6 and HHV-7/HCMV) were additionally identified. Interestingly, HHV-7 was only detected in the presence of other betaherpesviruses. Sequencing information confirmed betaherpesviruses infection. High hepatic enzyme levels and INR values>1.4 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 and serum samples were positive for some betaherpesviruses, suggesting an association with ALF. Coinfection of HHV-7 with HHV-6 or HCMV was additionally detected, suggesting that the precursor betaherpesviruses infection can trigger HHV-7 infection. Based on these results, we propose that ALF patients should be screened for the presence of betaherpesviruses.

Herpesviruses oral shedding and viremia in renal transplant recipients: A longitudinal study

“…In general, the limit of detection of the pan‐herpes test performed in our study were 10‐400 genome copies per reaction 9 . This sensitivity is comparable to qPCR real time protocols described previously for some herpesviruses and also the qPCR protocol used in our laboratory 25,26 …”

“…9 This sensitivity is comparable to qPCR real time protocols described previously for some herpesviruses and also the qPCR protocol used in our laboratory. 25,26 Our results showed that herpesviruses were co-detected in the participants' saliva before renal transplantation (32.8%) as well as 15-20 (56.1%) and 45-60 (52%) days after transplantation.…”

Presence and clinical impact of human herpesvirus‐6 infection in patients with moderate to critical coronavirus disease‐19

“…We present a retrospective study where hospitalized patients with signs and symptoms of moderate to severe COVID- The qPCR reaction for HHV-6 was performed using a primer and probe sequences for the U56 region of HHV-6. 17 To investigate the active infection, the HHV-6 detection was performed in the same samples used to detect SARS CoV-2, for this reason, the RNA extracted was used in this study. A saliva sample tested to intermediate replicative was used as positive control.…”

“…The qPCR reaction for HHV‐6 was performed using a primer and probe sequences for the U56 region of HHV‐6 17 . To investigate the active infection, the HHV‐6 detection was performed in the same samples used to detect SARS CoV‐2, for this reason, the RNA extracted was used in this study.…”

Longitudinal study on oral shedding of human betaherpesviruses 6 and 7 in renal transplant recipients reveals active replication

“…The quantitative real-time PCR assay was applied to measure viral loads [18] using U56 and U37 regions to target HHV-6 A/B and HHV-7, respectively. Primers and oligonucleotide TaqMan probes are presented in Table 1.…”