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As described earlier, we applied both multiplex PCR assays and culture-based tests for 52 environmental samples. In this regard, multiplex PCR assays showed that all of the Karaj River samples contained coliforms, E. coli, Shigella spp., Salmonella spp., and P. aeruginosa. However, only two out of 52 samples were contaminated with V. cholerae. Results of culture-based examinations justified the PCR assays, since exactly the same results were obtained, which confirms the excellent and ideal consistency and association between these methods (Table 4). Furthermore, for the spiked sample, all target genes were detected (Figure 2). These promising results proved the possibility of assessing six different types of bacteria in actual and artificial water matrices within just 16 hours (12 hours for enrichment and 4 hours for PCR assay).

Table 4

Number of samples and positive results

StrainNumber of samplesCP
Coliforms 52 52 52 
E. coli 52 52 52 
Shigella spp. 52 52 52 
Salmonella spp. 52 52 52 
P. aeruginosa 52 52 52 
V. cholerae 52 
StrainNumber of samplesCP
Coliforms 52 52 52 
E. coli 52 52 52 
Shigella spp. 52 52 52 
Salmonella spp. 52 52 52 
P. aeruginosa 52 52 52 
V. cholerae 52 

C: No. of positive results through culture-based examinations.

P: No. of positive results through PCR assays.

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