Fungi are a major agricultural, veterinary and medicinal problem across the world; Candid species are opportunistic infecting immunocompromised patients, ie those with AIDS, undergoing chemotherapy or transplant procedures. Aspergillus species affect human, animals and crops. Presently, lengthy culture methods are used clinically for microbe identification. Rapid diagnosis using molecular sequence recognition methods can be used to enable timely prescription of effective antimicrobial chemotherapy. Molecular methods while faster and more accurate are yet to have made an impact within the clinical setting, due to problems of specificity, complexity of process and insufficient reduction in time of screening and analysis. For molecular detection to work it must be specific, rapid and simple to use, in order to overcome the dominance of the present techniques. Presented here is a rapid, nanolitre sampling multiple probe approach developed for fungi Candida and Aspergillus species determination. The 18S ribosomal RNA small subunit sequence is present in high copy numbers per cell, allowing identification without fragment amplification. The variation within the molecule is limited therefore a bioinformatic analysis has been used to develop a multiple probe system to take advantage of conserved and variable regions. Samples are extracted rapidly by microwave, then passed through a micro channel device where by the sample is interrogated by multiple probes pattern on a surface, initially a reverse line blot membrane but now reduced to a microscope slide. The result is a method that provides rapid and accurate diagnosis of fungal pathogens.
Full conference title:
161st Society for General Microbiology
- SGM 161st (2007)