Department: Quality Assurance

DOC. No. : AV/MB/SOP/003 Page No. : Page 1 of 3 00

Effective Date: 01.06.06 Sub ect

Revision No. :

SOP for isolation ! i"entification of Salmonella spp.

Revision ( 00

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DOC. No. : AV/MB/SOP/003 Page No. : Page * of 3 00

Effective Date: 01.06.06 Sub ect

Revision No. :

SOP for isolation ! i"entification of Salmonella spp.

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Purpose +o ,efine proce,-re for Salmonella spp..isolation an, i,entification. Scope +/is proce,-re is applicable to isolate an, i,entif! Salmonella spp. Responsibilities Microbiologist AM.0-alit! Ass-rance Definition : SOP( Stan,ar, operating proce,-re. Proce"ure 1.1 Operation 1.1.1 1.1.* 1.1.3 +ransfer 1 gm or 1 ml of sample to 100 ml of n-trient brot/ in a sterile scre2.cappe, 3ar. S/a4e5 allo2 stan,ing for 6 /o-rs an, s/a4ing again. $oosen t/e cap an, inc-bate at 31.378# for *6 /o-rs.

/., Primary 0est 1.*.1 1.*.* 1.*.3 A,, 1.0 ml of t/e enric/ment c-lt-re to eac/ of t/e t2o t-bes containing A9 10 ml of Selenite. brot/ an, B9+etrat/ionate.bile.brilliant green brot/. %nc-bate bot/ t/e t-bes at 36.3:8# for 6: /o-rs. rom eac/ of t/ese t2o c-lt-res; s-bc-lt-re on at least t2o of t/e follo2ing fo-r agar me,ia. a9 Bism-t/ s-lp/ite agar b9 Brilliant green agar c9 'eo<!c/olate agar ,9 =!lose.l!sine.,eso<!c/olate agar %nc-bate t/e plates at 36.3:8# for 1:.*6 /o-rs. )pon e<amination; if none of t/e colonies conforms t/e ,escription as given in t/e table; t/e sample meets t/e re>-irements of t/e test for t/e absence of t/e gen-s Salmonella.

1.*.6 1.*.1

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DOC. No. : AV/MB/SOP/003 Page No. : Page 3 of 3 00

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Revision No. :

SOP for isolation ! i"entification of Salmonella spp.

0able: 0est for Salmonella 1e"ium #ismut$ sulp$ite agar #rilliant green agar DCA 23D Description of colony Blac4 or green Small; transparent an, colorless or opa>-e pin4is/ or 2/ite ?fre>-entl! s-rro-n,e, b! a pin4 or re, @one9 #olorless an, opa>-e; 2it/ or 2it/o-t blac4 centers. Re, 2it/ or 2it/o-t blac4 centers.

%f an! colonies conforming to t/e ,escription as given in t/e table; carr! o-t t/e secon,ar! test. /.- Secon"ary test 1.3.1 S-bc-lt-re an! colon! s/o2ing t/e c/aracteristics given in table in triple s-gar iron agar b! first inoc-lating t/e s-rface of t/e slope an, t/en ma4ing a stab c-lt-re 2it/ t/e same inoc-lating nee,le; an, at t/e same time inoc-late a t-be of -rea brot/. %nc-bate at 36.3:8# for 1:.*6 /o-rs. +/e formation of aci, an, gas in t/e stab c-lt-re ?2it/ or 2it/o-t concomitant blac4ening9 an, t/e absence of aci,it! from t/e s-rface gro2t/ in t/e +S% agar; toget/er 2it/ t/e absence of a re, colo-r in t/e -rea brot/; in,icates t/e presence of salmonella. %f aci, b-t no gas is pro,-ce, in t/e stab c-lt-re; t/e i,entit! of t/e organisms s/o-l, be confirme, b! aggl-tination tests.

1.3.* 1.3.3

1.3.6

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Documentation ( Microbiological test report

4.)

References: %P.A6/A.116/VO$..%%

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