Government health facilities were used as study sites. At each study site, a study physician conducted a physical examination, obtained written consent from participants’ parent and enrolled the eligible participants. Also, the physician administered a short questionnaire to collect demographic information and vaccination status. An experienced phlebotomist collected 1.0 ml blood by venipuncture.

In addition, as part of the study protocol, test for hemoglobin was conducted on infants and willing mothers through hemocue method and results shared immediately. Hemoglobin less than 11 gm/dL in infants and < 12 gm/dL among mothers was considered as anemia.

The blood samples for poliovirus antibodies were centrifuged every day, sera separated in cryovials and stored below −20 °C until shipped in dry ice to The Enterovirus Research Centre,

Mumbai on completion of enrolments. The samples were tested to determine neutralizing antibodies against all three poliovirus types using Sabin OPV strains in a modified micro- neutralization assay following a standard protocol [14]. Serial two-fold dilutions of test serum sample (1:8 to 1:1024) were reacted with 100 CCID50 of each of the three poliovirus types. HEp-2(C) cells were used to detect virus infectivity. All samples for polio antibodies were tested in triplicate beginning at a 1:8 dilution. Internal reference serum and virus back titration was included in each test run. Antibody titers were determined by Karber method. Positive controls were included in every run, to determine the antibody titers.

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