•Scientific questions: Expanding the Global Polio Eradication Initiative (GPEI) and establishing environmental surveillance systems in countries at risk of poliovirus transmission warrants the development of additional virus enrichment methods with reduced costs and shorter turnaround times.•Evidence before this study: The acute flaccid paralysis (AFP) case surveillance system is the gold standard for poliovirus surveillance; however, the “silent” transmission of poliovirus in the population makes environmental surveillance a crucial auxiliary tool for poliovirus surveillance.•New findings: The filtration method using a mixed cellulose ester (MCE) membrane is more sensitive than two-phase separation and can be used as a robust, sensitive, and cost-effective method to isolate enteroviruses from sewage.•Significance of the study: The filtration method using a mixed cellulose ester (MCE) membrane allows sensitive monitoring of the dynamic changes of enteroviruses in the environment. Additionally, this method enables researchers to track the actual dynamics of enterovirus transmission in human communities and provide early warnings for diseases in the population. This study aimed to compare the sensitivity of two-phase separation and the filtration method using a mixed cellulose ester (MCE) membrane to detect enteroviruses in sewage samples. From December 2015 to July 2016, four domestic sewage samples (1 L/sample) were collected monthly from the Guangzhou Liede Sewage Treatment Plant, and each sewage sample was divided into two aliquots (500 mL). The sewage sample was concentrated using the two-phase separation and the filtration method using an MCE membrane, and the treated solutions were inoculated into cells for enterovirus isolation. Polymerase chain reaction amplification, VP1 sequencing, and enterovirus molecular typing were performed on the positive isolates. The detection rates of poliovirus (PV) and non-polio enterovirus (NPEV) obtained using the filtration method using an MCE membrane were higher than those using the two-phase separation method. McNemar's test showed that the detection rates of PV, NPEV, type 1 Sabin-like (SL1), type 2 Sabin-like (SL2), and type 3 Sabin-like (SL3) strain were not statistically significant (P > 0.05). In Guangdong Province, China, the detection rates for PV and NPEV were 53.13% and 62.50% (20/32), respectively. Twenty-seven PVs were isolated, three highly variable strains of the type 1 vaccine, with seven nucleotide substitutions in the VP1 region, compared with the type 1 Sabin strains. Eighty-seven strains of NPEV were isolated and nine serotypes were identified, among which coxsackievirus B3 (CVB3), echovirus 6(E6), and echovirus 11(E11) were the dominant strains. The filtration method using an MCE membrane is more sensitive than two-phase separation and can be used as a robust, sensitive, and cost-effective method to isolate enteroviruses from sewage.