LAMP detection of Cryptococcus neoformans in AIDS patients

Subject Areas : Molecular Microbiology

Mohammad Hassan Shahhosseiny ¹ , Alireza Rezaei Amirabadi ² , Jalil Vand Yousefi ³ , Mohammad Ghahri ⁴ , Kayhan Azadmanesh ⁵ , Elham Moslemi ⁶

1 - Department of Microbiology, Islamic Azad University, Qods branch, Tehran, Iran
2 - Department of Microbiology, Islamic Azad University, Karaj branch, Alborz, Iran
3 - Department of Microbiology, Islamic Azad University, Karaj branch, Alborz, Iran
4 - Department of Biology, Imam Hossein University, Tehran, Iran
5 - Department of Virology, Pasteur Institute, Tehran, Iran
6 - Department of Biology, Islamic Azad University, Tehran east Branch, Tehran, Iran

Received: 2009-08-16 Accepted : 2010-01-15 Published : 2010-01-01

Keywords: LAMP, Cryptococcus neoformans, AIDS,

Abstract :

Background and Objective: Cryptococcus neoformans is opportunistic capsulated yeast that infects immunocompromised individuals. Infection occurs due to inhalation of the agent and common infection form is meningitis. Several methods are common to detect this life threthening agent but none of them have well accepted in sensitivity, specificity and rapidness. In this research the effort has been on installation and optimizing the LAMP assay in order to detect the agent itself and its distribution importance of Cryptococcus neoformans in serum specimens of AIDS patients. Material and methods: Atotal number of 34 serum specimens were collected from 34 AIDS patients. After applying sensitivity and specificity tests for the installed technique, optimization of the assay was performed on the specimens. The specimens got inactivated and after steps of DNA extraction, 6 designated primers for the target gene (ura5) were applyed for detection of Cryptococcus neoformans by using the LAMP method. Results: In this study 3 specimens of the total number of 34 AIDS patients were reported positive by using LAMP technique. This it reveals the importance of Cryptococcal infection in AIDS patients. Conclution: Loop-mediated isothermal amplification is an applicable, sensitive, quick and lucrative method for detection of Cryptococcus neoformans. Diagnostic laboratory can use this method without requiring any special instruments.

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