Vocma, Variation-tolerant Capture Multiplex analysis a new multiplex method for detecting virus, bacteria and fungi
Abstract number: P1825
Öhrmalm C., Eriksson R., Jobs M., Herrmann B., Melhus Å., Blomberg J.
Objectives: We have developed a new multiplex method for simultaneous detection of nucleic acid from virus, bacteria and fungi, called VOCMA, Variation-tolerant Capture Multiplex Analysis. Today the panels of VOCMA Gastroenterits and VOCMA Sepsis are 7-plex and 22-plex resp.
The method is variation tolerant i.e. it can detect nucleic acid from viruses like calici virus (winter vomiting disease) or influenza virus, which are highly disposed to genomic variation with the aim of avoiding the immune defense of the host. The variation tolerance is also allowing VOCMA to cover whole groups of similar genes like e.g antibiotic resistant genes as ctxm group I and ctxm group IV.
Methods: VOCMA is based on well established steps as nucleic acid extraction, followed by amplification and labelling of the target sequences of the pathogen. Target specific probes are attached to the different xMAP microspheres and a mixture of beads, consisting of the panel of interest, are allowed to hybridize to the sample. With a Luminex® 200 flow cytometer the amplified and labeled nucleic acid of the target genes are identified and quantified.
Result: The VOCMA method is a highly multiplex (e.g. 22-plex VOCMA sepsis) and provides tolerance to variation in the nucleic acid sequence. Even though it is variation tolerant the VOCMA is still highly specific for the intended target. The method, which is being further optimized, has shown an analytical sensitivity of 1010000 targets/reaction for both the VOCMA Gastroenterits and VOCMA Sepsis.
Conclusion: VOCMA is a multiplex and specific method for detection of nucleic acid of different microorganism. By utilizing xMAP technology and Luminex® 200 the composition of the multiplex panels can easily be changed, combined and expanded, which creates a large flexibility of the assay. A third panel which contains virus, bacteria and fungi causing respiratory infectious diseases is also under development. A multiplex method like VOCMA both saves time, reduces the cost per microbe analysed, as well as the consumption of sample, and increases the possibility of discovering co-infections.
|Session name:||Abstracts 20th European Congress of Clinical Microbiology and Infectious Diseases|
|Location:||Vienna, Austria, 10 - 13 April 2010|
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