A total of 237 representative V. cholerae O1 strains isolated between 2001 and 2012 were included for the analysis of rtxA, rstB, and gyrA genes. Additionally another 50 strains isolated between 1989 and 2000 were also included in the genotyping of gyrA. All the V. cholerae strains used for this study were selected from the strain repository of National Institute of Cholera and Enteric Diseases (NICED), Kolkata, India and were isolated from the hospitalized cholera patients. NICED Human Ethical committee had approved the study (Approval No. C-48/2012-T&E). All the participants provided their written consent to participate in this study and these documents were maintained in the Clinical Division of NICED. The content and consent procedure was also approved by the above committee. The strains were grown in Luria Bertani broth (Becton Dickinson, Sparks, MD, USA) for 18 hrs. and then streaked on Luria agar (Becton Dickinson, Sparks) plates. Identity of these strains was reconfirmed serologically by the slide agglutination with O1 specific polyclonal antiserum and serotype specific antisera (Becton Dickinson, Sparks). V. cholerae O1 strains EL-1786 (Ogawa), N16961 (Inaba) and 0395 (Ogawa) were used as standard strains for the Haitian, El Tor and classical type, respectively.
Luria agar
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Luria agar is a solid growth medium commonly used for the cultivation and isolation of bacteria, particularly Escherichia coli (E. coli). It provides essential nutrients and growth factors required for the proliferation of bacterial cultures.
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2 protocols using luria agar
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Genotypic analysis of Vibrio cholerae O1
2
Characterization of V. cholerae O1 Strains
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V. cholerae O1 strains isolated in Delhi between 2004 and 2012 was characterized in this study. 14 strains from 2004, 13 strains from 2005, 20 from 2006, 39 from 2007, 23 from 2008, 13 from 2009, 11 from 2010, 15 from 2011 and 27 strains from 2012 were considered. V. cholerae strains examined for this study were selected from the strain repository of Maharishi Valmiki Infectious Diseases Hospital, Delhi, India and were isolated from the hospitalized cholera patients. After incubation for 18h in Luria Bertani broth (Becton Dickinson, Sparks, MD, USA), all the strains were then streaked on Luria agar (Becton Dickinson) plates.
Serology of these strains was confirmed by the slide agglutination with O1 specific polyvalent antiserum and serotype specific antisera (Becton Dickinson) . In this study, V. cholerae O1 strains EL-1786 (Ogawa), N16961 (Inaba) and O395 (Ogawa) were used as standard strains for the Haitian, El Tor and classical types, respectively.
Serology of these strains was confirmed by the slide agglutination with O1 specific polyvalent antiserum and serotype specific antisera (Becton Dickinson) . In this study, V. cholerae O1 strains EL-1786 (Ogawa), N16961 (Inaba) and O395 (Ogawa) were used as standard strains for the Haitian, El Tor and classical types, respectively.
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