Mycoplasma infection and rheumatoid arthritis. Analysis of their relationship using immunoblotting and an ultrasensitive polymerase chain reaction detection method

Abstract

Objective. To examine the relationship between infection with Mycoplasma and the development of rheumatoid arthritis (RA) and juvenile rheumatoid arthritis ( J U ) . Methods.Immunoblotting of patient synovial fluid and sera on detergent-phase membrane protein extracts of various Mycoplasma species was carried out to learn whether patients exhibited serologic evidence of previous exposure to mycoplasmas. Moreover, an ultrasensitive polymerase chain reaction (PCR) method was developed for assessing whether Mycoplasma DNA could be detected in synovial fluid from patients and controls.Results. Immunoblotting provided serologic evidence of previous Mycoplasma exposure in patients and controls. The genus-specific PCR detected known human Mycoplasma species and could reliably detect <5 copies of Mycoplasma hominis, Mycoplasma fermentans, or a molecular mimic control in synovial fluid. Repeat testing revealed no evidence of Mycoplasma DNA in patient synovial samples.Conclusion. This study provided serologic evidence suggesting that, while previous exposure to Mycoplasma was common, there was no detectable persistence of Mycoplasma DNA in the synovial fluid or tissue of patients with RA or JRA.

Phylogeny-Based Rapid Identification of Mycoplasmas and Ureaplasmas from Urethritis Patients

“…Species-specific PCR has proven successful in achieving rapid detection, but it requires several sets of primers, since several species of mycoplasmas or ureaplasmas have been implicated as the etiologic agents of urethritis. In contrast, Hoffman et al (10) reported the use of a primer set, GPO-1 and MGSO, for detecting not only human mycoplasmas but also ureaplasmas. However, their PCR was not sensitive enough to detect low amounts of mycoplasmas or ureaplasmas in clinical samples.…”

Structural Analysis of TCRα and β Chains from Human T‐Cell Clones Specific for Small Nuclear Ribonucleoprotein Polypeptides Sm‐D, Sm‐B and U1–70 kDa: TCR Complementarity Determining Region 3 Usage Appears Highly Conserved

“…Amplification of a portion of the TCRAC region was performed in parallel as a positive control. Extensive precautions were taken to ensure that cross-contamination of samples did not occur [21]. The amplified DNA was subjected to gel electrophoresis in 3% NuSieve, 1% Sea Kem agarose (FMC, Rockland, ME, USA) and stained with ethidium bromide.…”

Site-Specific Proteolysis of the MALP-404 Lipoprotein Determines the Release of a Soluble Selective Lipoprotein-Associated Motif-Containing Fragment and Alteration of the Surface Phenotype of Mycoplasma fermentans

“…Whether RF is produced, or further processed, in the human host is unknown, as is its precise function. Nevertheless, human serum Ab to MALP-404 has been noted previously (19). This product may now be more fully explored by retrieval of soluble, authentic RF product from in vitro culture.…”