Development of a supersensitive polymerase chain reaction method for human T lymphotropic virus type II (HTLV-II) and detection of HTLV-II proviral DNA from blood donors in Japan

Abstract

A supersensitive polymerase chain reaction procedure was developed to detect human T-lymphotropic virus type II (HTLV-II) proviral genome. Six primer pairs covering the various regions of HTLV-II were compared and selected on the basis of specificity and sensitivity. Among them, one primer pair of the pol region of HTLV-II (II pol) was able to amplify and detect even 0.1 fg of the cloned plasmid HTLV-II DNA (seven copies) by regular ethidium bromide staining on polyacrylamide gel. By using this procedure, we screened 189 HTLV-I seropositive blood donors from Yamaguchi and Fukuoka Red Cross Blood Centers, Japan. There were four positive samples detectable with the HTLV-II-specific pol primer pair, as well as with the HTLV-I tax primer pair. The amplified DNAs of two specimens were cloned and sequenced. The sequences of the HTLV-I tax region from both specimens were identical to that of HTLV-I. On the other hand, those of the HTLV-II pol region were identical to that of HTLV-II, except for one base substitution in a clone from one subject. These results indicate that dual infection of HTLV-I and HTLV-II in the same persons occurs among Japanese blood donors.

Letters to the Editor

“…Levels of potassium in the control arm (y-irradiated only) were comparable with those previously found by Rivet et al [2] who noted levels of 30 mMat 4 days and Ramirez et al [3] who reported 68 mM K+ at 14 days. Walther-Wenker et al [4] made similar findings and now transfuse babies with BPF-4-filtered SAG-M product, irradiated and transfused at any time up to 14 days.…”

“…First, not all transfusions of bacterially contaminated blood components cause fever [2], Second, when fevers occur in septic transfusion reactions, temperature eleva tions may overlap the range of fevers from nonseptic reactions [2,3]. Third, febrile platelet transfusion reactions are caused frequently by inflammatory cytokines released from leukocytes during storage [5], or by alloimmune reactions to leukocyte antigens [6], reducing the specificity of fever as an indicator of a septic platelet transfusion reac tion.…”

“…Both viruses can be transmitted by blood cell components, whereas plasma or sera are not infectious [3,4], Although a high proportion of recipients of contam inated HTLV-positive blood become infected [5], HTLV-I-related ill nesses develop in only a few of them [6], The transmission of HTLV-I I by blood transfusion is less efficient, and its pathogenicity is still not well known [7].…”

“…Georgetown University Medical Center, Washington, D.C., USA As the number of reports of sepsis caused by transfusion of con taminated platelet concentrates increases, estimates of the risk of this potentially fatal complication are updated accordingly [1][2][3], A recent study of 32,000 random-donor platelet concentrates prepared at a large Canadian Red Cross blood center revealed that approximately 1 in 1,000 units was contaminated by bacteria [4], The current risk of sig nificant bacterial contamination of a platelet concentrate exceeds the combined risk of transfusion-transmitted infection by human immu nodeficiency viruses (HIV-1/2), human T-lymphotropic viruses (HTLV-I/II), hepatitis B virus, and hepatitis C virus [4].…”

“…All 19 units were then y-irradiated to a dosage of 34.5 Gy delivered over 7 min. All units were sampled before and after fil tration (if applicable) and irradiation, followed by sampling at 3,7,14,21,28,35 and 42 days. The supernatant was removed and stored at -40°C until assayed for Na+, K+ and glucose concentration.…”