More recently, the use of multiplex PCR has provided
a rapid and inexpensive method for detection of several
pathogens in a single tube. Multiplex PCR assay is capable of screening various microbial organisms simultaneously
or identifying different alleles of one organism. Multiplex
PCR uses multiple pairs of primers to amplify multiple
targets in a single reaction tube and requires no additional
procedures and equipment. After first reported in 1988
(ref. 11), this has became a fast and simple method for
clinical and research laboratories. In multiplex PCR, the
primers and conditions that are applicable in a monoplex
setting no longer produce the same results because the
primers for different organisms interfere with each other
and reduce the sensitivity. Therefore, optimization of
conditions and concentration of reagents used need to be
standardized, keeping in view the ‘nature’ of each primer
as well as requirement of sensitivity in that particular
situation.