Quantification of anaerobic ammonium-oxidizing bacteria in enrichmentcultures by quantitative competitive PCR
HAO Chun , WANG Huan , LIU Qinhua , LI Xudong
DOI:
Received December 23, 2008,Revised April 17, 2009, Accepted , Available online
Volume 21,2009,Pages 1557-1561
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The anaerobic ammonium-oxidizing (ANAMMOX) bacteria were enriched from a sequencing batch biofilm reactor (SBBR). A
quantitative competitive polymerase chain reaction (QC-PCR) system was successfully developed to detect and quantify ANAMMOX
bacteria in environmental samples. For QC-PCR system, PCR primer sets targeting 16S ribosomal RNA genes of ANAMMOX bacteria
were designed and used. The quantification range of this system was 4 orders of magnitude, from 103 to 106 copies per PCR,
corresponding to the detection limit of 300 target copies per mL. A 312-bp internal standard was constructed, which showed very
similar amplification e ciency with the target amxC fragment (349 bp) over 4 orders of magnitude (103–106). The linear regressions
were obtained with R2 of 0.9824 for 103 copies, 0.9882 for 104 copies, 0.9857 for 105 copies and 0.9899 for 106 copies, respectively.
Using this method, ANAMMOX bacteria were quantified in a shortcut nitrification/denitrification-anammox system which was set for
piggery wastewater treatment.
