Single-stranded conformation polymorphism of large subunit of ribosomal RNA is best suited to diagnosing fungal infections and differentiating fungi at species level.
In the last decade, various polymerase chain reaction (PCR)-based methods have been developed using ribosomal RNA (rRNA) for the identification of medically important fungi. In the present study, large subunit (LSU) and small subunit (SSU) of fungal rRNA were amplified and analyzed by single-stranded conformation polymorphism (SSCP) of nested PCR, restriction digestion, and SSCP of digested products. The relationship between several clinical isolates of patients suffering from aspergillosis, candidiasis, cryptococcosis, keratitis, and skin and nail infections has been established with standard fungal cultures using the SSU- and LSU-specific primers. Single-stranded conformation polymorphism of restriction profile of amplified products of LSU-specific primers was successfully used to differentiate fungi up to genus and species level.
Kumar
Manish
M
Fermentation Technology Division, Medical Mycology Laboratory, Central Drug Research Institute, Lucknow-226 001, India.
Shukla
Praveen Kumar
PK
D002965:Classification / Q000379:methods; D017931:DNA Primers; D003937:Diagnosis, Differential; D005658:Fungi / Q000145:classification* / Q000235:genetics*; D006801:Humans; D009181:Mycoses / Q000175:diagnosis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D018807:Polymorphism, Single-Stranded Conformational; D012331:RNA, Fungal / Q000032:analysis; D012335:RNA, Ribosomal / Q000145:classification / Q000235:genetics*; D012016:Reference Values
D017931:DNA Primers; D012331:RNA, Fungal; D012335:RNA, Ribosomal