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

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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