Establishment and evaluation of SYBR Green fluorescent PCR for detection of Aeromonas

ObjectiveTo establish a SYBR Green fluorescent PCR assay for the detection of Aeromonas.

MethodsSYBR Green fluorescent PCR was established by using 16S rRNA gene amplification primers of Aeromonas reported, and its specificity, sensitivity and practical application value were evaluated.

ResultsThe detection limit of SYBR Green fluorescent PCR was 10^–7 ng/ml. The assay to detect 21 Aeromonas strains showed positive results, but the assay showed negative results for Vibrio, Escherichia coli, Salmonella, Yersinia enterocolitica and Shigella. For 104 stool samples of healthy people, Aeromonas was isolated from 8.65% of the samples, 28.85% of the samples showed positive results in SYBR Green fluorescent PCR. Eight of them were positive by both methods. If the pathogen was isolated after positive SYBR Green fluorescent PCR detection, one sample (11.11%) would be missed, but 71.15% (74/104) of the workload of isolation, culture and identification might be reduced.

ConclusionSYBR Green fluorescent PCR has high specificity and sensitivity. It can reduce the workload by about 70% in the early screening of large samples, and is suitable for the monitoring of Aeromonas infection and carrier.