University of Medicine and Dentistry of New Jersey

Yong Liu

Michael J Leibowitz

The present invention pertains to an in vitro method for assaying for an inhibitor of the catalytic Group I self-splicing intron reaction in the nuclear rRNA genes of Pheumocystis carinii which comprises the steps of (a) providing a DNA template containing the intron (I) from the 26S rRNA gene in Pneumocystis carinii and a portion of the 5' and 3' flanking exons (E1 and E2, respectively) between nucleotides 1963 and 2267 of 26S rRNA (660 nucleotides of amplified rRNA gene including the group I intron); (b) preparing an RNA precursor by transcription of the DNA template in the presence of labeled nucleoside triphosphates to produce a labeled RNA precursor (E1-I-E2); (c) purifying the RNA precursor; (d) incubating the RNA precursor and the inhibitor in the presence of guanosine triphosphate and magnesium ions; and (e) determining the degree of inhibition by the inhibitor on the intron splicing reaction in the RNA precursor by measuring the amount of labeled splicing intermediates and splicing products.

In vitro assay for inhibitors of the intron self-splicing reaction in

The present invention pertains to a method for diagnosing for Pneumocystis carinii by detecting the presence of a nucleic acid sequence containing the 26S rRNA gene specific for Pneumocystis carinii. More particularly, this invention relates to a method for diagnosing for Pneumocystis carinii which comprises amplifying a sample of DNA from Pneumocystis carinii by polymerase chain reaction (PCR) using species specific primers and detecting the PCR products with species specific radioactive or non-radioactive oligonucleotide probes. This invention also relates to a method for diagnosing for various species of Pneumocystis carinii by detecting the presence of a nucleic acid sequence containing the particular 16S or 26S rRNA gene sequence specific for that species of Pneumocystis carinii.

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Piscataway, NJ, United States of America

Yong Liu