A reverse transcription-nested PCR assay for HHV-6 mRNA early transcript detection after transplantation.
Monitoring of human herpesvirus-6 (HHV-6) reactivation is important, especially in immunocompromised patients such as transplant recipients. Reverse transcription PCR (RT-PCR) is a useful method to distinguish between latent and active infection. Here, a RT-nested PCR coupled with a colorimetric plate hybridization assay was established to detect HHV-6 types A and B U79/80 mRNAs. After confirming the reliability of the assay on HHV-6 cultures, it was applied to the detection of HHV-6 reactivation after renal (27 patients), bone marrow (14 patients) or heart (7 patients) transplantation. A total of 206 blood samples were tested from renal (137), bone marrow (58) and heart (11) transplant recipients. U79/80 mRNAs were found in 32 samples that were considered as indicative of HHV-6 reactivation: 15, 13 and 5 from kidney, bone marrow and heart transplant recipients, respectively. Finally, U79/80 mRNA detection was correlated with clinical manifestations including leucopenia, skin rash, graft rejection or dysfunction and diarrhoea.
Pradeau
Karine
K
Laboratoire de Virologie, CHU Dupuytren, Limoges, France.
Bordessoule
Dominique
D
Szelag
Jean-Christophe
JC
Rolle
Florence
F
Ferrat
Pierre
P
Le Meur
Yann
Y
Turlure
Pascal
P
Denis
François
F
Ranger-Rogez
Sylvie
S
D000328:Adult; D000368:Aged; D000818:Animals; D016026:Bone Marrow Transplantation / Q000009:adverse effects; D002460:Cell Line; D017931:DNA Primers; D005260:Female; D005814:Genes, Viral / Q000235:genetics; D016027:Heart Transplantation / Q000009:adverse effects; D015654:Herpesvirus 6, Human / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D016030:Kidney Transplantation / Q000009:adverse effects; D008297:Male; D008875:Middle Aged; D011183:Postoperative Complications / Q000175:diagnosis / Q000821:virology; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D019349:Roseolovirus Infections / Q000175:diagnosis* / Q000209:etiology / Q000821:virology; D012680:Sensitivity and Specificity; D014775:Virus Activation
D017931:DNA Primers; D012333:RNA, Messenger; D012367:RNA, Viral