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Detection of Mycoplasma genus & Mycoplasma fermentans by PCR in patients with Chronic Fatigue Syndrome (CFS)

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By Vojdani A, Choppa PC, Tagle C, Andrin R, Samimi B, Lapp CW • • December 15, 1998

Mycoplasma fermentans and other Mycoplasma species are
colonizers of human mucosal surfaces and may be associated
with human immunodeficiency virus infection. While many
infectious agents have been described in different percentages
of patients with Chronic Fatigue Syndrome (CFS), little is
known about the prevalence of mycoplasmas and especially M.
fermentans in CFS patients. A polymerase chain reaction
(PCR)-based assay was used to detect Mycoplasma genus and M.
fermentans genomes in peripheral blood mononuclear cells
(PBMC) of CFS patients. Blood was collected from 100 patients
with CFS and 50 control subjects. The amplified products of
717 bp of Mycoplasma genus, and 206 bp of M. fermentans were
detected in DNA purified from blood samples in 52% and 34% of
CFS samples, respectively. In contrast, these genomes were
found in only 14% and 8% of healthy control subjects
respectively (P < 0.0001). All samples were confirmed by
Southern blot with a specific probe based on internal
sequences of the expected amplification product. Several
samples, which were positive for Mycoplasma genus, were
negative for M. fermentans indicating that other Mycoplasma
species are involved. A quantitative PCR was developed to
determine the number of M. fermentans genome copies present in
1 microg of DNA for controls and CFS patients. Mycoplasma copy
numbers ranging from 130 to 880 and from 264 to 2400 were
detected in controls and CFS positive subjects, respectively.
An enzyme immunoassay was applied for the detection of
antibodies against p29 surface lipoprotein of M. fermentans to
determine the relationship between M. fermentans genome copy
numbers and antibody levels. Individuals with high genome copy
numbers exhibited higher IgG and IgM antibodies against M.
fermentans specific peptides. Isolation of this organism by
culture from clinical specimens is needed in order to
demonstrate specificity of signal detected by PCR in this

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