TaqMan RT-PCR and VERSANT HIV-1 RNA 3.0 (bDNA) assay Quantification of HIV-1 RNA viral load in breast milk.

BACKGROUND: Transmission of HIV via breast milk is a primary cause of pediatric HIV infection in developing countries. Reliable methods to detect breast milk viral load are important. OBJECTIVE: To correlate the ability of the VERSANT HIV 3.0 (bDNA) assay to real-time (RT) TaqMan PCR in quantifying breast milk HIV-1 RNA. STUDY DESIGN: Forty-six breast milk samples that had been spiked with cell-free HIV-1 and eight samples spiked with cell-associated HIV-1 were assayed for HIV-1 RNA by both VERSANT HIV 3.0 and TaqMan RNA assays. RESULTS: Only assays on the cell-free samples were statistically compared. Both a Deming regression slope and a Bland-Altman slope indicated a linear relationship between the two assays. TaqMan quantitations were on average 2.6 times higher than those of HIV 3.0. A linear relationship was observed between serial dilutions of spiked cell-free HIV-1 and both the VERSANT HIV 3.0 and the TaqMan RNA assays. CONCLUSION: The two methods correlated well although the VERSANT HIV 3.0 research protocol quantified HIV-1 RNA slightly lower than TaqMan.

Testicular volume: comparison of orchidometer and US measurements in dogs.

PURPOSE: To compare the accuracy and precision of orchidometer and ultrasonographic (US) measurements of testicular volume in a canine model. MATERIALS AND METHODS: Volume measurements of 18 canine testes were obtained by using Prader and Rochester orchidometers. Testes were scanned with two linear-array US transducers with imaging frequencies of 6-13 MHz and 5-10 MHz. For each transducer, testicular volumes were calculated by using three formulas: length (L) x width (W) x height (H) x 0.52, L x W(2) x 0.52, and L x W x H x 0.71. Testes were weighed following bilateral orchiectomies. True testicular volume was determined by using the formula volume = weight/density. Paired t tests were used to assess whether mean measurement biases differed significantly from zero. The relationship between true and measured volume was evaluated with a linear regression model. RESULTS: US volume measurements demonstrated lower variability and better linear fit compared with orchidometry (R(2) = 0.75-0.90 vs R(2) = 0.14-0.38). The formula L x W x H x 0.71 had the smallest mean bias relative to true volume with use of either transducer over the entire volume range. CONCLUSION: US methods of testicular volume measurement are more accurate and precise than orchidometry. The formula L x W x H x 0.71 provides a superior estimate of testicular volume and should be used in clinical practice.