Generation of HIV-1 and Internal Control Transcripts as Standards for an In-House Quantitative Competitive RT-PCR Assay to Determine HIV-1 Viral Load

Table 3

Evaluation of the presence of specific RNA (RT-PCR signal) and the DNA contamination (PCR signal) of the HIV-1 and IC transcripts.

Name	Dilution	Average RT-PCR fluorescence value (FU) ± SD	Average PCR fluorescence value (FU) ± SD
HIV-1-negative control	—	4.05±0.09	3.56±0.18
HIV-1-positive control (RT-PCR)	—	181.9±2.83	—
HIV-1-positive control (PCR)	—	—	117.80±5.36
IC-negative control	—	3.37±0.11	3.35±0.08
IC-positive control (PCR)	—	—	141.81±2.14
T-HIV1-2^b	1 : 4 × 10^3	165.92±2.28	172.55±8.15
	1 : 4 × 10^6	181.67±4.72	31.93±0.17
	1 : 4 × 10^7	187.14±7.57	3.87±0.24
	1 : 4 × 10^8	170.72±8.42	3.87±0.03
	1 : 4 × 10^9	171.24±5.59	4.65±0.27
T-IC-2^b	1 : 4 × 10^3	210.00±0	3.26±0.06
	1 : 4 × 10^6	107.55±0.36	3.35±0.04
	1 : 4 × 10^7	11.87±0.38	3.29±0.01
	1 : 4 × 10^8	3.43±0	3.23±0.11
	1 : 4 × 10^9	3.46±0.11	3.43±0.03

^bTranscript treated with 4 u of RNase-free DNase per 1 μg of DNA used in the in vitro transcription.
FU: fluorescence units; SD: standard deviation; IC: internal control.