Use of real-time PCR to monitor human herpesvirus 6 reactivation after allogeneic bone marrow transplantation.
Reactivation of human herpesvirus 6 (HHV-6) is common following allogeneic marrow transplantation, however, little is known about the immune control and pathogenic potential of HHV-6 infection after transplantation. In order to determine whether reactivation of HHV-6 plays an important role in the development of complications in patients undergoing allogeneic bone marrow transplantation or not, we developed a very rapid quantification of viral DNA using a LightCycler. The amount of viral DNA was determined using a supernatant of a chronically infected cell line [TaY(OK)] which contains a known amount of viral DNA. Peripheral blood cells were collected from 5 patients undergoing allogeneic bone marrow transplantation once before transplant and once per week after transplant for 8-24 weeks. The real-time PCR system revealed that there was a linear correlation in the range of 101 to 105 molecules of reference. Using this system, we have found the presence of non-diagnosed HHV-6 reactivation as well as symptomatic infection, indicating the potential for routine implementation of this technology for laboratory diagnosis of HHV-6 infection. Our study shows that this method of rapid quantification of HHV-6 genomes by the real-time PCR using a LightCycler may be useful not only to understand the reconstitution of the immune system following marrow transplantation but also to manage the care of patients.
Ohyashiki
J H
JH
Department of Virology, Medical Research Institute, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8510, Japan. junko@hh.iij4u.or.jp
Suzuki
A
A
Aritaki
K
K
Nagate
A
A
Shoji
N
N
Ohyashiki
K
K
Ojima
T
T
Abe
K
K
Yamamoto
K
K
D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D016026:Bone Marrow Transplantation; D004247:DNA / Q000097:blood / Q000235:genetics; D004279:DNA, Viral / Q000097:blood / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006566:Herpesviridae Infections / Q000821:virology*; D015654:Herpesvirus 6, Human / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D014184:Transplantation, Homologous; D014775:Virus Activation
D004279:DNA, Viral; D004247:DNA