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Mycoplasma fermentans is a likely causative agent of HIV-associated nephropathy. In a pilot study, M. fermentans DNA was detected with polymerase chain reaction (PCR) in urine samples from renal allograft recipients; nine (39.1%) out of 23 renal allograft recipients (most of whom had chronic allograft rejection) and none of the 20 controls, were infected with M. fermentans. A cross-sectional study was conducted to investigate the prevalence of M. fermentans in urine samples from renal allograft recipients. Midstream urine samples were centrifuged at 13,000 x g, purified with QIAamp and tested with PCR using RW004/RW005 and an internal control to screen for the presence of inhibitors. Of the 264 participants recruited, 263 completed the questionnaire (172 men, 92 women); 53 had chronic renal allograft rejection, 106 had chronic renal dysfunction without rejection, 69 had a normal renal allograft for more than 3 months and 35 had a renal allograft for less than 3 months. All urine samples yielded positive results for the internal control. Mycoplasma fermentans DNA was detected once i prospectively collected urine samples. The only individual infected with M. fermentans was also seropositive for HIV-1. This study demonstrates that M. fermentans can be at most sporadically detected in urine from patients living with a renal allograft but is not implicated in chronic rejection of allograft.

The polymerase chain reaction (PCR) is a highly sensitive technique for the detection of lymphoma cells presenting specific rearrangements or translocations, like the t(14;18). However, few methods are available for the quantitative evaluation of clinical samples, especially using nested PCR. In this study, we describe a method for the semiquantitative estimation of low numbers of lymphoma cells using the nested PCR methodology. Samples evaluated consisted of RL cells, a non-Hodgkin's lymphoma cell line harbouring a bcl-2 translocation, mixed with normal mononuclear cells (MC) obtained from bone marrow or peripheral blood. DNA was extracted from samples with RL:MC ratios ranging from 5 x 10(-2) to 5 x 10(-7), and amplified for detection of this translocation. The product of this first amplification reaction was serially diluted 10-fold from 10(0) to 10(-7). Then each fraction was PCR-amplified with nested oligonucleotide primers. Aliquots of the second amplification were electrophoresed on a 2% agarose gel and stained with ethidium bromide. Within the range tested, there was a log-linear relationship between the number of PCR positive bands and the number of translocated cells in each sample. This procedure was highly reproducible in experiments evaluating the interrun and intrarun variability. In addition, there was no interaction with normal cells obtained from peripheral blood or bone marrow, or from different individuals. Furthermore, results were identical with two different cell lines and consistent with patient lymphoma cells obtained from a lymph node biopsy.(ABSTRACT TRUNCATED AT 250 WORDS)