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Background: In the last two decades, cryptococcosis has been gaining a distinct public
health importance due to the growing number of AIDS
cases. Considering the low sensitivity of direct examination with India ink and
culture, use of sensitive techniques is crucial in the diagnosis of cryptococcal
meningitis. Polymerase Chain Reaction (PCR)
can be used to directly detect Cryptococcus neoformans in CSF
samples to increase the diagnostic power in cases where conventional
methods are unable to detect the organism.
Methods : In this cross-sectional study, CSF samples were
obtained from 25 patients suspected of
having neurocryptococcosis. The patients were referred to the Medical Mycology
Laboratory of the School of Public Health affiliated to Tehran University of
Medical Sciences from March 2009 to February 2010.
Three different methods, direct India ink examination, culture and PCR
were used to evaluate the CSF samples. Two 102
and 106 of Cryptococcus neoformans dilutions in
1ml of CSF
were prepared and examined by the three methods. In PCR
method, two primer pairs were selected to amplify the Cryptococcus
neoformans URA5 gene. The sequences
of primers were for A, B, C and D
serotypes.
Results : Only in one case PCR, as well as direct
examination and culture were positive. All the other samples were negative in PCR,
direct examination or culture. Both CSF dilutions
were positive in the three tests in the mentioned patient and the positive
control.
Conclusion: PCR
method can efficiently identify both control and positive samples of Cryptococcus
neoformans.
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