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SOP for i!entification an! isolation of P.aeruginosa.

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SOP for i!entification an! isolation of P.aeruginosa. ).( Purpose ,o -efine proce-.re for isolation of Pse.-omonas aer.ginosa / its i-entification. Scope ,0is proce-.re is applicable to isolate an- i-entif" P.aer.ginosa. Responsibilities Microbiologist AM12.alit" Definition : SOP) Stan-ar- operating proce-.re. Proce!ure ..) ,ransfer t0e 3.antit" of t0e pretreate- preparation 410 gm or 10 ml sample 5 60 ml b.ffer p0osp0ate 7.+ p89: e;amine- containing 1gm or 1ml of t0e pro-.ct to 100 ml of fl.i- so"abean casein -igest me-i.m. ..+ Mi; an- inc.bate at 3<137=$ for +414> 0o.rs. .., ';amine t0e me-i.m for gro?t0 an- if gro?t0 is present: strea@ a portion of t0e me-i.m on t0e s.rface of cetrimi-e agar me-i.m: eac0 plate- on Petri -is0es. ..- $o er an- inc.bate at 3<137=$ for 1>1+4 0o.rs. ... &f .pon e;amination: none of t0e plates contains colonies 0a ing t0e c0aracteristics liste- in t0e table of t0e me-ia .se-: t0e sample meets t0e re3.irement for free-om from P.aer.ginosa. ..* &f an" colone" conforming to t0e -escription in table are pro-.ce-: carr" o.t t0e o;i-ase an- pigment tests. ../ Strea@ representati e s.spect colonies from t0e agar s.rface of cetrimi-e agar me-i.m on t0e s.rface of Pse.-omonas agar me-i.m for t0e -etection of fl.orescein an- pse.-omonas agar me-i.m for -etection of p"oc"anin containe- in Petri -is0es. ..0 $o er an- in ert t0e inoc.late- me-ia an- inc.bate at 33137=$ for not less t0an 3 -a"s. ..1 ';amine t0e strea@e- s.rfaces .n-er .ltra1 iolet lig0t. ';amine t0e plates to -etermine ?0et0er colonies conforming to t0e -escription gi en in t0e table.

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SOP for i!entification an! isolation of P.aeruginosa. ..)( &f gro?t0 of s.spect colonies occ.rs: place + or 3 -rops of a fres0l" prepare- 1A ?/ sol.tion of B:B:B1:B1:1tetra1met0"l141 p0en"lene-iamine -i0"-roc0lori-e on filter paper an- smear ?it0 t0e colon" or transfer colonies to strips or -is@s of filter paper t0at pre io.sl" 0as been impregnate- ?it0 B:B1-imet0"l1p1 p0en"lene-iamine -i0"-roc0lori-e if t0ere is no -e elopment of a pin@ colo.r :c0anging to p.rple: t0e sample meets t0e re3.irement of t0e test for t0e absence of Pse.-omonas aer.ginosa. 2able: 2ests for P. aeruginosa

3e!ium Cetrimi!e agar Pseu!omonas agar me!ium for !etection of fluorescein Pseu!omonas agar me!ium for !etect ion of pyocyanin *.(

C#aracteristic 4luorescence in colonial 5% lig#t morp#ology Cenerall" greenis0 Creenis0 Cenerall" colo.rless to "ello?is0 Dello?is0

O6i!ase test Positi e Positi e

7ram stain Begati e ro-s Begati e ro-s

Cenerall" greenis0 Bl.e

Positi e

Begati e ro-s

Documentation : Microbiological test report References: &P166/A1116/VO%.1&&

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