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Indian Journal of Medical Microbiology
Medknow Publications on behalf of Indian Association of Medical Microbiology
ISSN: 0255-0857
EISSN: 0255-0857
Vol. 29, No. 4, 2011, pp. 406-410
Bioline Code: mb11097
Full paper language: English
Document type: Research Article
Document available free of charge

Indian Journal of Medical Microbiology, Vol. 29, No. 4, 2011, pp. 406-410

 en Standardization of fungal polymerase chain reaction for the early diagnosis of invasive fungal infection
Deshpande, P; Shetty, A; Mehta, A; Kapadia, F; Hedge, A; Soman, R & Rodrigues, C

Abstract

Background: An early initiation of antifungal therapy in invasive fungal infections (IFIs) is critical in reducing the high mortality rate. Current diagnosis of fungal infection relies on microscopy, culture, antigen, antibody specific tests and histological diagnosis. However, these tests either lack sensitivity or specificity. There is thus the need for a rapid, specific and accurate diagnostic method.
Objective: The aim of our study was to establish PCR for the rapid detection of Candida check for this species in other resources and Aspergillus check for this species in other resources species in clinical specimens with improved sensitivity and specificity.
Materials and Methods: A total of 71 proven cases of IFI (confirmed by culture) were collected. A total of 15 healthy, 15 patients suffering from bacterial sepsis and 15 patients with HIV, HBV viral infections were included as controls. Clinical specimens were subjected to a standardized nested amplification to produce Round I (504 bp) and Round II (150 bp) amplicons. Restriction digestion was performed on these products for further identification.
Results: Analytical sensitivity was determined using 10 6 -10 CFU/ml of cell suspension. The lower detection limit of the assay was 10 CFU/ml of blood. This test was 100% sensitive and specific with a positive predictive value of 100% and a negative predictive value of 96.7%.
Conclusion: The assay was found to be effective for the rapid detection of Candida and Aspergillus in clinical specimens.

Keywords
Aspergillus, Candida, nested polymerase chain reaction, polymerase chain reaction, restriction enzyme digestion

 
© Copyright 2011 Indian Journal of Medical Microbiology.
Alternative site location: http://www.ijmm.org

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