Table 2.

Efficacy of DNase treatment of cellular RNA extracted from cellular specimens

StepNo. of HIV-1 RNA copies/PCR mixture for the following specimens with the indicated no. of DNA copiesa:
Amplicor RNA diluentControl, 20,000 DNA copiesb,cPatient AdPatient B, no DNAdPatient C, no DNAd
200 DNA copiese20,000 DNA copiese200 DNA copiescNo DNA
1. No processing20642,730NDf NDNDNDND
2. RNeasy protocol without DNase INDND70,503NDNDNDND
3. Standard protocol with 1× DNase INDND94965135
4. Extended protocol with 2× DNase INDND<2516696
No. of samplesg 1122211
  • a 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.

  • b PBMCs from an HIV-negative donor.

  • c DNA was added after step 2.

  • d PBMCs from HIV-positive donors receiving HAART.

  • e DNA was added after step 1.

  • f ND, not determined.