7/27/2019 Av-mb-sop-001 (Bis) Sop for Isolation & Identification of s.aureus

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Department: Quality Assurance DOC. No. : AV/MB/SOP/001

Page No. : Page 1 of 3

Effective Date: 01.06.06 evision No. : 00

!u"#ect !OP for isolation $ i%entification of !tap&. Aureus.

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evision + evise% Pages Description Date00 Original issue 01.06.06

7/27/2019 Av-mb-sop-001 (Bis) Sop for Isolation & Identification of s.aureus

2/3

Department: Quality Assurance DOC. No. : AV/MB/SOP/001

Page No. : Page ( of 3

Effective Date: 01.06.06 evision No. : 00

!u"#ect !OP for isolation $ i%entification of !tap&. Aureus.

1.0 Purpose

)o *efine proce*+re for Stap,ylococc+s a+re+s-isolation an*

i*entification.

,.0 !cope

),is proce*+re is applicable to isolate an* i*entify Stap,ylococc+s

a+re+s.

-.0 esponsi"ilities

Microbiologist AM-+ality Ass+rance

.0 Definition :

SOP& Stan*ar* operating proce*+re.

/.0 Proce%ure

/.1 #noc+late sample on Bair*-Parer me*i+m an* in salt me*i+m.

/., #nc+bate t,e Bair*-Parer me*i+m at 32! for at least 30 ,o+rs.

/.- From t,e salt me*i+m mae s+bc+lt+res on t,e Bair*-Parer

me*i+m an* inc+bate at 32! for 30 ,o+rs./. $4amine not less t,an 5 of t,e s+spect colonies of Stap,ylococc+s

on t,e soli* me*ia an* mar o+t as many s+spect colonies as

possible to investigate.

/./ est for i%entification

/./.1 ram2s stain

),e stain consists of

a30.5 6 met,yl violet or crystal violet in 7ater.

"3#o*ine sol+tion816 io*ine an* ( 6 potassi+m io*i*e

in 7ater9. c3 !o+nter stain 80.1g ne+tral re* 0.(ml of 16 acetic

aci* an* 100 ml of 7ater9.

/./., On a clean grease-free sli*e very lig,t an* t,in smear

covering a small area is ma*e *irectly from li:+i* c+lt+re

an* in clean tap 7ater if from soli* me*ia.

/./.- ),e smear is fi4e* by passing to an* fro over a flame an*

coole*.

/./. !over t,e smear 7it, t,e stain 8a9 for 30 secon*s.

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7/27/2019 Av-mb-sop-001 (Bis) Sop for Isolation & Identification of s.aureus

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Department: Quality Assurance DOC. No. : AV/MB/SOP/001

Page No. : Page 3 of 3

Effective Date: 01.06.06 evision No. : 00

!u"#ect !OP for isolation $ i%entification of !tap&. Aureus.

/././ Po+r off t,e stain an* 7as, 7it, 8b9 an* t,en cover 7it,8b9 an* allo7 remaining for 30 secon*s.

/./.6 ;as, off 7it, et,anol +ntil t,e *ye ceases to stream o+t.

;as, in r+nning tap 7ater an* apply8c9 for abo+t one

min+te. ;as, in tap 7ater an* *ry for e4amination.

/.6 Coagulate est& ),is test may be carrie* o+t by t,e follo7ing

met,o*s&

a3 !li%e met&o% < $m+lsify a portion of t,e s+spect

colony in normal saline or 7ater. Mi4 t,is 7it, a

straig,t 7ire *ippe* in ,+man or rabbit plasma.

!oag+lase-positive stap,ylococci pro*+ce visiblecl+mping imme*iately.

"3 u"e met&o% < $m+lsify a single s+spect colony

from a (= ,o+r gro7t, of bloo* agar me*i+m in 1 ml

citrate* rabbit plasma *il+te* 1 in 5 in 0.>5 6 saline.

),e rest is +s+ally carrie* o+t in narro7 t+bes. Place

in an inc+bator or preferably in 7ater-bat, at 32!.

Observe every ,o+r to note clotting of plasma.

Rea*ing s,o+l* be carrie* o+t for as long as possible

preferably avoi*ing overnig,t inc+bation. Positive

control 7it, a no7n coag+lase-positive strain of

Stap,ylococc+s an* a control of t,e *il+te* plasma7it,o+t inoc+l+ms s,o+l* be incl+*e* in t,e test

6.0 Documentation:

Microbiological )est Report

4.0 eferences:#S& 5>> 8Part ##9 < 1?@

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