TABLE 4.
Cell-associated viral copies in low-level infection subjects: multiple replicate nested nef-p15 multiplex PCRs
| Baby | Tissue | Time point (wk) | DNA per reaction (ng) | No. of replicates | No. positive | Total DNA tested (μg) | Estimated copy no./μg DNAb | Estimated copy no./million cellsb,c |
|---|---|---|---|---|---|---|---|---|
| B2 | PBMC | 4 | 200 | 5 | 0 | 1 | <2.0 | <13.2 |
| PBMC | 24 | 200 | 5 | 0 | 1 | <2.0 | <13.2 | |
| Tonsil | 24 | 200 | 20 | 0 | 4 | <0.5 | <3.0 | |
| Ileum | 24 | 200 | 20 | 0 | 4 | <0.5 | <3.0 | |
| Spleen | 24 | 200 | 20 | 0 | 4 | <0.5 | <3.0 | |
| Thymus | 24 | 200 | 20 | 0 | 4 | <0.5 | <3.0 | |
| B7 | PBMC | 4 | 200 | 10 | 0 | 2 | <1.0 | <6.0 |
| B9 | PBMC | 24 | 200 | 10 | 0 | 2 | <1.0 | <6.0 |
a
Limit of detection of nested nef-p15 PCR assay: 2 to 5 copies/μg DNA. The limit of detection for the nested multiplex PCR was estimated by performing multiple independent limiting-dilution nested nef-p15 PCRs on a standard panel of templates that contained 1, 10, 100, and 1,000 copies/PCR.
b
The copy number per microgram of DNA was estimated using Quality Software (24).
c
The estimated copy number per microgram DNA was extrapolated to the copy number per million cells based on the amount of DNA per cell (6.6 pg DNA per cell).