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 6

Accuracy of the qRT-PCR assay by using HIV-1 controls and the transcripts as standards.

Sample	HIV-1 RNA (IU/mL)		Log10 expected results/actual results
	Expected results	Actual results^c
IS (1 : 100)	3.63𝐸+03^a	3.81𝐸+03	−0.02
IS (1 : 10)	3.63𝐸+04^a	4.81𝐸+04	−0.12
IS	3.63𝐸+05^a	5.78𝐸+05	−0.20
HIV1-235	1.00𝐸+06^b	2.33𝐸+06	−0.37
HIV1-157	1.30𝐸+06^b	1.03𝐸+06	0.10

^aThe value for each IS sample was obtained from dilutions made to the HIV-1 international standard (code 97/650) that contained 363 078.05 IU/mL (reconstituted in 1 mL of ultrapure water) as instructed by the NIBSC manual.
^bThe value for each material was determined by the NucliSens EasyQ HIV-1 v1.1 assay (bioMerieux, Boxtel, The Netherlands).
^cHIV-1 viral loads estimated from the in-house qRT-PCR assay using the HIV-1 (T-HIV1-2^b) and IC (T-IC-2^b) transcripts as standards.