Ocena wystepowania zakazen Pneumocystis badaniem wymazow z gardla i jamy ustnej metoda nested PCR

Badano materiał uzyskany z wymazów z gardła i jamy ustnej metodą nested PCR w celu wykrycia infekcji Pneumocystis u ludzi i szczurów laboratoryjnych. Stwierdzono, że czułość nPCR umożliwia wykorzystanie wymazów do badania rozprzestrzenienia pneumocystozy wśród ludzi i zwierząt laboratoryjnych oraz stwarza możliwość bezpiecznej diagnostyki infekcji u pacjentów, których stan kliniczny uniemożliwia uzyskanie materiału z dolnych dróg oddechowych.

A method of collecting pharyngeal and oral swabs from humans and laboratory rats, to be examined later for Pneumocystis infection with simple and nested PCR, was developed. The swabs were obtained from 15 HIV-infected patients, including 5 suffering from PCP, and from 10 immunocompetent children aged 2 to 6 years. Furthermore, the swabs were taken from 30 healthy laboratory rats and 23 animals subjected to immunosuppressive treatment. DNA of Pneumocystis was detected in all the examined rats with nPCR method, but only in 47% of healthy animals when simple PCR was used. Nested PCR examination of swabs collected from human subjects revealed the infection with Pneumocystis in all HIV-infected patients with PCP, and in 8 out of 10 symptomless carriers of Pneumocystis; moreover, the examination was positive in 2 out of 10 immunocompetent children. It was concluded, that noninvasive method of collecting pharyngeal and oral swabs in conjunction with very sensitive method of amplification DNA by nPCR is suitable for measuring the prevalence of Pneumocystis infection in the populations of humans and laboratory animals. The developed method offers a possibility of safe diagnosis of pneumocystosis in patients whose clinical status precludes collection of BAL through bronchoscopy.