Reverse transcriptase viral load correlates with RNA in SIV/SHIV-infected macaques.
Monitoring of viral load in macaques has usually been carried out using in-house PCR-based methods. A novel viral load (VL) kit (ExaVir Load) based on the measurement of lentivirus reverse transcriptase (RT) activity provides a potential alternative to methods that measure plasma viral RNA. RT is a fundamental and conserved activity of all retroviruses and the method should theoretically detect RT from all lentiviruses. To test this we compared VL measured by a commercially available RT kit with an in-house QC RT-PCR in macaques infected with SIV and SHIV. Both RT and RNA levels were measured over time in both sets of macaques. Results indicated that the relationship between both tests was strong for SIV and SHIV (r = 0.95 and r = 0.92, p < 0.0001, respectively). The VL trends also followed each other, indicating that both techniques measured the same process of viral replication. Furthermore, the RT load obtained using standardized control plasma samples supplied by NIBSC gave values close to the designated VL. However, when comparing RT load with QC RT-PCR a consistently three to five time higher level was obtained with the RT assay, highlighting potential differences in assay calibration. Even so, the data suggest that the RT assay is both sensitive and robust for use in the SIV/SHIV macaque model, particularly where molecular-based assays for SIV VL determinations are not easily available. The assay is also a commercially available kit and hence has the potential to reduce the variability seen between laboratories using in-house PCR.
Corrigan
Gary E
GE
Swedish Institute for Infectious Disease Control, Solna, Sweden., Microbiology Tumorbiology Center, Karolinska Institute, Stockholm, Sweden.
Hansson
Eva Olausson
EO
Mörner
Andreas
A
Berry
Neil
N
Källander
Clas F R
CF
Thorstensson
Rigmor
R
D000818:Animals; D005260:Female; D016086:Lentivirus / Q000201:enzymology; D008251:Macaca / Q000821:virology*; D012367:RNA, Viral / Q000097:blood*; D012194:RNA-Directed DNA Polymerase / Q000097:blood* / Q000378:metabolism; D011933:Reagent Kits, Diagnostic / Q000821:virology*; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000302:isolation & purification*; D019562:Viral Load
D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D012194:RNA-Directed DNA Polymerase