Selective detection of Mycoplasma fermentans using the polymerase chain reaction (PCR).

Important note: Information in this article was accurate in 1992. The state of the art may have changed since the publication date.

Selective detection of Mycoplasma fermentans using the polymerase chain reaction (PCR).

Abstr Annu Meet Am Soc Microbiol. 1991 May 5-9;91:134 (abstract no. G-5). Unique Identifier : AIDSLINE ASM91/0030292
Wang R; Hu W; Dawson MS; Lo SC; Shih JW; ARP, Armed Forces Institute of Pathology, Washington, DC.

Abstract: We previously developed a PCR assay for M. fermentans (incognitus strain). The assay using primer pairs RS47 and RS49 however failed to detect many mycoplasmas of the M. fermentans species, including PG-18, the prototype of M. fermentans and new clinical isolates from patients with AIDS. Recently, we reported the presence of a 1.4 kb insertion sequence (IS)-like element in multiple copies in the M. fermentans genome. A new assay using the PCR procedure for amplifying a 205 nt specific gene sequence within the IS-like element combined with sequence-specific oligonucleotide probe hybridization has been developed. The assay using primer pairs, RW004 (24 nt) and RW005 (24 nt), and Tag enzyme, selectively amplifies DNA from M. fermentans mycoplasmas. In contrast, DNA from other species of human mycoplasmas, common tissue culture contaminating mycoplasmas, and bacteria, as well as human, monkey and mouse tissues do not produce the amplified DNA products which are verified by oligonucleotide probe RW006 representing a segment of the intervening sequences in the IS-like element between RW004 and RW005. The assay can detect a single microorganism in each reaction. This highly sensitive and specific assay should prove to be valuable in detection of AIDS-associated mycoplasma infections.

Keywords: Acquired Immunodeficiency Syndrome/MICROBIOLOGY DNA, Bacterial/ANALYSIS Gene Amplification Genes, Bacterial Human Mycoplasma fermentans/GENETICS/*ISOLATION & PURIF Oligonucleotide Probes Polymerase Chain Reaction ABSTRACT
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