Article

Multiplex PCR for the Detection of Mycoplasma fermentans, M. hominis, and M. penetrans in Patients with Chronic Fatigue Syndrome, Fibromyalgia, Rheumatoid Arthritis, and Gulf War Syndrome

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  • immunosciences lab
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Abstract

Summary A multiplex polymerase chain reaction (PCR) was used to detect mycoplasma infection in human DNA samples of patients with CFS and related illnesses. One set of oligonucleotide primers which are specific for a highly conserved region among all members of the genus Mycoplasma along with three other primer sets which are specific for Mycoplasma fermentans, M. hominis, and M. penetrans species were used in this assay. The sensitivity of detection was determined by adding known mycoplasma DNA copy numbers to 1 μg of genomic DNA from healthy subjects. Each sample was subjected to 40 cycles of amplification. The detection level was determined to be 7, 7, 9, and 15 mycoplasma DNA copies per μg of human genomic DNA for M. genus, M. fermentans, M. hominis, and M. penetrans, respectively. The assay was applied to DNA extracted from the PBMCs of individuals suffering from chronic fatigue syndrome (CFS) (n = 100), fibromyalgia (FMS) (n = 40), rheumatoid arthritis (RA) (n = 60), and gulf war syndrome (GWS) (n = 60) and compared to age- and sex-matched healthy individuals (n = 160). The percentage of M. genus infection detected in CFS, FMS, RA, and GWS was 52,54, 49, and 55%, respectively. M. fermentans was detected in 32, 35, 23, and 36%, M. hominis was detected in 9, 8,11, and 5%, and M. penetrans was detected in 6, 4, 7, and 3% of CFS, FMS, RA, and GWS patients, respectively. M. genus, M. fermentans, M. hominis, and M. penetrans were detected in 15, 8, 3, and 2% of healthy matched controls. This assay provides a rapid and cost efficient procedure to screen clinical samples for the presence of three potentially pathogenic species of Mycoplasma with a high level of sensitivity and specificity.

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... Jones and Clark (102), exercise physiologists, have developed guidelines for exercise programs for fibromyalgia syndrome. As up to 75% of ME/CFS patients meet the criteria for FMS (49) this program would also be appropriate for ME/CFS. We have summarized Jones' and Clark's suggestions and adapted them for ME/CFS. ...
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Bacterial and viral infections are purported to be associated with several fatigue illnesses, including Chronic Fatigue Syndrome (CFS), Fibromyalgia Syndrome (FMS), Gulf War Illnesses (GWI) and Rheumatoid Arthritis (RA), as causative agents, cofactors or opportunistic infections. We and others have looked for the presence of invasive pathogenic mycoplasmal infections in patients with CFS, FMS, GWI and RA and have found significantly more mycoplasmal infections in CFS, FMS, GWI and RA patients than in healthy controls. Most patients had multiple mycoplasmal infections (more than one species). Patients with chronic fatigue as a major sign often have different clinical diagnoses but display overlapping signs/symptoms similar to many of those found in CFS/FMS. When a chronic fatigue illness, such as GWI, spreads to immediate family members, they present with similar signs/symptoms and mycoplasmal infections. CFS/FMS/GWI patients with mycoplasmal infections generally respond to particular antibiotics (doxycycline, minocycline, ciprofloxacin, azithromycin and clarithromycin), and their long-term administration plus nutritional support, immune enhancement and other supplements appear to be necessary for recovery. Examination of the efficacy of antibiotics in recovery of chronic illness patients reveals that the majority of myco-plasma-positive patients respond and many eventually recover. Other chronic infections, such as viral infections, may also be involved in various chronic fatigue illnesses with or without mycoplasmal and other bacterial infections, and these multiple infections could be important in causing patient morbidity and difficulties in treating these illnesses.
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