TABLE 2.
Step | No. of HIV-1 RNA copies/PCR mixture for the following specimens with the indicated no. of DNA copiesa:
|
||||||
---|---|---|---|---|---|---|---|
Amplicor RNA diluent
|
Control, 20,000 DNA copiesbc | Patient Ad
|
Patient B, no DNAd | Patient C, no DNAd | |||
200 DNA copiese | 20,000 DNA copiese | 200 DNA copiesc | No DNA | ||||
1. No processing | 206 | 42,730 | NDf | ND | ND | ND | ND |
2. RNeasy protocol without DNase I | ND | ND | 70,503 | ND | ND | ND | ND |
3. Standard protocol with 1× DNase I | ND | ND | 9 | 49 | 65 | 13 | 5 |
4. Extended protocol with 2× DNase I | ND | ND | <2 | 51 | 66 | 9 | 6 |
No. of samplesg | 1 | 1 | 2 | 2 | 2 | 1 | 1 |
Copies of restriction enzyme (PvuII)-digested diluted plasmid pBH10 (5); calculation included the consideration that one molecule of double-stranded DNA represents two templates for the PCR.
PBMCs from an HIV-negative donor.
DNA was added after step 2.
PBMCs from HIV-positive donors receiving HAART.
DNA was added after step 1.
ND, not determined.