TY - JOUR
T1 - Quantification of human T-cell lymphotropic virus type 1 proviral load by quantitative competitive polymerase chain reaction
AU - Albrecht, Björn
AU - Collins, Nathaniel D.
AU - Newbound, Garret C.
AU - Ratner, Lee
AU - Lairmore, Michael D.
N1 - Funding Information:
The authors would like to thank Yasuko Rikihisha for providing the Bio Photonics gel documentation system, Patrick L. Green for helpful discussion and critical reading of the manuscript, and Tim Vojt for preparation of the Figures. This work was supported in part by National Institute of Health grants NCI CA55185, NIAID AI40302 and NIAID AI01474.
PY - 1998/11/15
Y1 - 1998/11/15
N2 - The polymerase chain reaction (PCR) has been established as a highly sensitive technique for detection of viral DNA or RNA. However, due to inherent limitations of PCR the amount of amplified product often does not correlate with the initial amount of template DNA. This is particularly true for PCR detection of viral infections that are characterized by low in vivo viral copy numbers in certain stages of the infection, such as human T-cell lymphotropic virus type 1 (HTLV-1) and simian T-cell lymphotropic virus type 1 (STLV-1). Therefore, we developed a quantitative competitive polymerase chain reaction (qcPCR) for detection of HTLV-1 and STLV-1 proviral DNA. The assay was optimized using an infectious HTLV-1 clone, ACH, HTLV-1 infected cell lines, MT-2.6 and HUT-102 and STLV-1 infected lines Kia and Matsu. Applicability of this system was demonstrated by determining HTLV-1 proviral load in peripheral blood mononuclear cells (PBMC) of human subjects with HTLV-1 associated diseases and an asymptomatic carrier as well as rabbits infected experimentally. This qcPCR method, the first designed specifically for HTLV-1 and STLV-1, will provide an important tool for pathogenesis studies of HTLV-1 and for evaluating the efficacy of antiviral drugs and vaccines against the viral infection using animal models. Copyright (C) 1998 Elsevier Science Ireland Ltd.
AB - The polymerase chain reaction (PCR) has been established as a highly sensitive technique for detection of viral DNA or RNA. However, due to inherent limitations of PCR the amount of amplified product often does not correlate with the initial amount of template DNA. This is particularly true for PCR detection of viral infections that are characterized by low in vivo viral copy numbers in certain stages of the infection, such as human T-cell lymphotropic virus type 1 (HTLV-1) and simian T-cell lymphotropic virus type 1 (STLV-1). Therefore, we developed a quantitative competitive polymerase chain reaction (qcPCR) for detection of HTLV-1 and STLV-1 proviral DNA. The assay was optimized using an infectious HTLV-1 clone, ACH, HTLV-1 infected cell lines, MT-2.6 and HUT-102 and STLV-1 infected lines Kia and Matsu. Applicability of this system was demonstrated by determining HTLV-1 proviral load in peripheral blood mononuclear cells (PBMC) of human subjects with HTLV-1 associated diseases and an asymptomatic carrier as well as rabbits infected experimentally. This qcPCR method, the first designed specifically for HTLV-1 and STLV-1, will provide an important tool for pathogenesis studies of HTLV-1 and for evaluating the efficacy of antiviral drugs and vaccines against the viral infection using animal models. Copyright (C) 1998 Elsevier Science Ireland Ltd.
KW - Animal model
KW - Diagnosis
KW - HTLV-1
KW - Pathogenesis
KW - Quantitative competitive polymerase chain reaction
KW - Retrovirus
UR - http://www.scopus.com/inward/record.url?scp=0031727523&partnerID=8YFLogxK
U2 - 10.1016/S0166-0934(98)00087-1
DO - 10.1016/S0166-0934(98)00087-1
M3 - Article
C2 - 9870588
AN - SCOPUS:0031727523
VL - 75
SP - 123
EP - 140
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 2
ER -