Quantitative competitive-polymerase chain reaction
Quantitative competitive PCR (QC-PCR) is a rapid quantitative
method, which involves the coamplification of a target
DNA sample with known amounts of a competitor DNA
having nucleotide sequence similar to the target. QC-PCR
thus allows the determination of absolute number of target
molecules in comparison to the amount of competitor DNA.
Precision of this method exceeds that of other quantitative
Appl Microbiol Biotechnol (2012) 95:1135–1154 1145PCR procedures including RT-PCR. The whole procedure
requires low-cost equipments and consumables
and can be carried out in <24 h. Reilly and Attwood
(1998) quantitatively measured Clostridium proteoclasticum
from cow rumen. Subsequently, Koike and
Kobayashi (2001) used this approach to enumerate F.
succinogenes, R. albus, and R. flavefaciens in sheep
rumen. Their data indicated that the minimum detection
levels were 1–10 cells in pure cultures and 103–4 cells/
ml in mixed cultures. In another study, Reilly et al.
(2002) quantified proteolytic bacteria viz. Streptococcus
spp., Butyrivibrio fibrisolvens, P. bryantii, Eubacterium
spp., and Prevotella spp. in cow rumen. Similarly, QCPCR
was applied to the study the influence of a host’s
diet on Butyrivibrio and Pseudobutyrivibrio populations
(Mrazek et al. 2006). Sekhavati et al. (2009) had applied
this technique to the estimation of fungal populations
in Holstein steers.
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