Iso 6579-1 2017 [PDF]

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INTERNATIONAL STANDARD



ISO



6579-1 First~dition Z017-O~



Microbiology of the food chain Horizontal method for the detection, enumeration and serotyping of Salmonella Part 1: Detection of Salmonella spp. Microhi()ICJ9"~de /(1 chatlle uJ;,"entairl! - Mcthotit!hQri%olfto/~ ptJur la ~herche.



Ie dt!llombremettt et Ie s~rol)'p(Jge des



Salrn(lflella PQrrJ~ 1:Rt!ch~rcht! da Sahnon~I1l'ls.DP.



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"ISO l017



ISO6579·t:2017(E)



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All rfghi. re....r.rwed. Unleis odJel'Wtse SJlKified, no pJt1. of this p-ubUcatinn IO"Ybe reproduced or-I.ltilited otMn'll$e In any fonn fir by any ,uean,s. elrttf'ollic or nN!'chanbl. htdudlog photOcopying..or postlng on the Internet or an Intrall,et..lwlth(lur prior\vrilU:l(prnl'lission ..PU'luission cal' be requested from eftber ISO ilt tbc! address below ,o,.-ISCYs menl\l.er body in the country o( the n!'q(IHI:tI~ 150 CftP)'l'lghl nlrtC.



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ISO6S7~·1::Z017(E)



Contents



Scope,_



2



NonnatIVe references __



3



1 Prlnclple .._ ...._ ._ .._. _.. . _. __ ...._ .... ~ .._ .... _ .... _ .... 2 4.a Cen"'t'aJ_,,_._ ,,'_" ~ ~ ~~., .2 4.2 Pre·.nrlchqll:Jlt In I1lIn·~dcai"u IIqulUmedlulll ......_ .... _._ ...._ ._.. , .....2 43 Enrichment in/on $.el~ctiVetnedi~, 4.4 ·~I.tlnc \lut ~Itij ,1111\1h, Itt> "'P"'"mI'dt rm 'I hail I'f.mmltli,e, Intfir'll:'tll"""1 or~~llIll!IUlI$'lWV"1'I111Ielu:.. 141,J lioUIrgtl'Y.llllIl'IJ,ItlJI,H,I,,'U;UIl '~IIIIISO • .,l~) noKr. I'll CIII rhc \lllIrC. Il\t'll'llilltburall'l' dQllely wllh 'lit .. h,l'Io"",.rtm~1 1i1""ITt.II",,'tlnIQlI !:'III'IImISldnn (IEel"" ..II "mil.", at 'i:!~cr(l'ts, V,ll f .I.e 1lI1Mlj'fialI.ll, tit. ~?1fcrIlIllI!IJllIU"'" O"Jrurr.. ""..w,t1 tw: trl'l' C&ftI,.,."i'I'V'pr.i!'" ISO J'lEllnll'hll! >1lI,r iii111_ Iii 0 lifo"" :,,!on 6/" trI,' I -2110;;



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ISO 6579-1:2017(8)



7 SaropUng Salllpling',s not part ollh.



Inotb01tJt1K'ei(!ed ill Ihlu(oCluncnt (see "''' specific !lIt.l·natlOnal Sl;'Indal1"i d."lillg wi! It lh~ 1)!odU',."mP'l!rnedj. If th~t~ Is 110s"odn~'nt.m"ti"JI~1SlilIldllnl. it is n:(ilS\ lbe m1lWlll.eor p,oQltngon Ihe.



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ISO6579-t:20t7(E)



9.5.4



Serologicaltesting



9.5.4.1 General The pure colonies (9.5..2) showing typical biochemical reactions for Salmonella (9..5.3) are also tested for the presence of 5{Jlmon""o 0- and H·nntlgens (and, In arras where Salmolldlu Typhl Is expected III the food supply. also for VI-antigen) by slide agglutination using polyvalent antisera (.IL..I!). Tile pure colonies are cultured on a non-selective ag3r medium (B..2) and tested for auto·agglutlnation. Strains that are autc-agglutlnable cannot be tested for the presence of Salmonelk: antigens. Use the antisera according to the manufncturer's Instructions If different from the method described belowte detect the presence of Salmonella O·and H-antlgens (and If necessary, also for Vl-antlgen]. The following tests ~to~)



arc the mlnlmullI required for serological testing ofSlIlmOllel/o spp.



Further guidance on serologlcal confirmation and on serotyping is given in ISO/TR 6579·31l!1. Elimination of auto·.agglutfnable stralns



9.5.4.2



Place one drop of saline solution (lUJ) on a clean gl.ss slide. Using a loop, disperse part ofthe colony to be tested In the saline 10 obtaln a homogeneous ond turbid suspension. Rode the slide gently for 5 s to 60 s (depending on the manufacturer's instructions). Observe the suspension. preferably against a dark background.Hrhe bacteria have formed granules IIIthe suspenslon, this indicates auto·agglutination and serological confirmsnen will become complicated. Additional information on the treatment of'auto-agglurtnattng stratns can be found in ISO/TR 6579·31211). 9.5.4.3



BxomlnnUon for O-anUgens



Using on. nen-auto-agglunnatlng pure colony, proceed according to~ auti-O sera (B.t4) in place of the sallne sclurion.



using one drop of polyvalcm



If agglutination occurs, this Is considered a pesltlve reactton. 9.S.4.4



Exruninatlon forVI·antigens



(optional) using one drop of anTI-VI



Using one no,,-auto·aggluth18tlng pure colony, proceed according 10 ~ scm (lUi) in place orthe saline solutlon. If agglutin~tion occurs, this is considered n po~itivC'reaction. 9.5.4.5



Examination for O-ontlgcns



Using one non·auto-agglutinAting pure colony, proceed according to.9..S.i..Zusing one drop of polyvalent anti-H sera (~) in place of the saline solution. If agglutination occurs. this is considered a positive reaction. 9.5.5



Interpretanon ofblocbemlcal



und serological ne"CUeDS



Iahl£ 2 gives the Interpretation of the confirmatory tests used~.



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ISO 6579·1:2017(E)



Table 2 -Interpretation Biochemical reactions Typical



Auto·aggtutination No



orconfinnatory



tests



Serological reactions O-.nd 1I'"II.lge..



Interpretation Slrllns c('Insld



data t;d.nbof nfump.I"" I·.tamed o(ter ellllluadllllll(lru. data Test por.tjon 1;i~e. ill g



Spc7" 32 5



STM44' 32



5E91'



19



32 5 19



95



95



9S



10



10



10



5 19



dare Test portion size, in g Spcctlklty.ln % Sensitivity per serevar and level, in % LOOseper serovnr (95 'l(, con(!dMcelnterv.I). du/test portion 1.001;4 ~"~1I0' STMSH' 33 33 33



Parameter Number of ,,"rtklp.tlng collabor"tors Number of 5aIJlp1e~per-collaborator Number of conaboraters ret>Jncd amr evoluAtlon of the-data Number or 5Aluples retalned arter cvaluildon of the data 're:lststIII tho recovery of 501111011.1/0 1ypbl olld 5a//II/)/w}/a Paratyph).



D.6 Confirmation 0.6.1



Geneml



See2.S. 0.6.2



Interpretation of the biochemical tests



Sa/moll./Io l'Yphi and Salmo/HIlla Pannyphl generally show the reaeuens given in Tahle J.



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ISO6S79-1:Z017(E)



AnnexE (informative)



Examples of selective plating-out media NOTti1 Derivedfl'omIl.f.r~ncc llD.l. Rel""duced by permission oflhelloy.1 So< ..



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>
rizolltal method for the detection eJlumerotiQII ana SJII'otypi',g of Sa 1111Oil gila -



Pao-t3: Guld.lin"" for scrolyping or 5011110110/10 spp,



(251



ISO 18593, Microbiology ol food and onlmalleedl"l/ stllffs - Horitontal methods for somplilIjJ techniques Jrom surfoces USingCOIIUlelplotesand swabs



[261



ISO/TS 17728, Microbiology of the rood chain - Sampllll9 {lUhniques for microbiological ollolysis (J{/'oud alllt fl!'TERS. Enhanced culture techniques far the detection or Salmonella. Vet.Roc. 2001, pp. 539-540



[34J



COMMUNITY REFllRENCE LABORATORYFOR SIILMONIiLM. Newsletter 200B. Vol. 14, no. 2, page 8. Available at: http;llww\y eurlsalolonrlla ru/dsresollrce1.type:pdf&dh;·posU·jon-:lnJjnc& a)Jj