© Liofilchem - ChromaticTM ESBL and ChromaticTM ESBL+AmpC...

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Chromatic TM ESBL ENGLISH Chromatic TM ESBL+AmpC Chromogenic media for detection of ESBLs and AmpC in Enterobacteriaceae directly from clinical specimens. DESCRIPTION Chromatic TM ESBL is a chromogenic medium used for detection of Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. ESBLs are enzymes that hydrolyze most penicillins and cephalosporins, inhibited by β-lactamase inhibitors such as clavulanic acid, sulbactam and tazobactam. Chromatic TM ESBL+AmpC is a chromogenic medium used for detection of ESBL and AmpC producers. AmpC-type enzymes hydrolyze penicillins, cephalosporins and monobactam and are poorly inhibited by the classical ESBL inhibitors, especially clavulanic acid. ESBL-producing Enterobacteriaceae have become one of the most important causes of nosocomial community-acquired infections caused by Escherichia coli and Klebsiella pneumoniae, but also other Gram-negative bacteria. Patients infected by ESBL producers often received inadequate empirical therapy until the pathogen’s resistance is recognized. Early detection is, therefore, essential in order to limit the spread of these pathogens. Organisms producing enough AmpC β-lactamase will typically give a positive ESBL screening test but fail the confirmatory test involving increased sensitivity with clavulanic acid. TYPICAL FORMULA (g/l) Peptone Mix 43.2 Chromogenic Mix 1.0 Agar 15.0 Selective Mix 0.5 Final pH 7.2 ± 0.2 at 25°C METHOD PRINCIPLE Peptones supply amino acids, nitrogen, carbon, minerals, vitamins and other nutrients which support the growth of microorganisms. Chromogenic mix allows the identification of microorganisms on the basis of the colony color and morphology. Agar is the solidifying agent. The Selective mix in Chromatic™ ESBL inhibits the ESBL-non-producing organisms, including AmpC producers. The Selective mix in Chromatic™ ESBL+AmpC inhibits the ESBL-non-producing organisms, but allows the growth of AmpC producers. PREPARATION Dehydrated medium Suspend 29.6 g of the powder in 500 milliliters of distilled or deionized water. Mix well. Heat until completely dissolved. Sterilize in autoclave at 121°C for 15 minutes. Cool to 45-50°C. Aseptically, add 5 ml previously reconstituted supplement, mix well and dispense in Petri dishes. Supplements Add 5 ml distilled or deionized water to the lyophilized supplement tube to be used. Mix well using a vortex mixer before adding to the medium being prepared. TEST PROCEDURE Inoculate the plates by streaking directly the specimen onto the agar surface. Incubate aerobically at 37°C for 18-24 hours. INTERPRETING RESULTS After incubation observe the color of the colonies and interpret the results as indicated in the ID table. ID Table. Microorganism* Typical colony color E. coli Pink-reddish-mauve Klebsiella spp, Enterobacter spp, Serratia spp Green-blue Proteus spp Brown *On Chromatic TM ESBL medium only ESBL-producing organisms grow, while Chromatic TM ESBL+AmpC allows the growth of both ESBL and AmpC producers. See pictures in Appendix I. APPEARANCE Dehydrated medium: fine, dry, homogeneous, free of extraneous material, beige. Lyophilized supplements: whitish button, limpid solution once reconstituted. Prepared medium: slightly opalescent, amber. STORAGE The powder is very hygroscopic, store the powder at 10-30°C, in a dry environment, in its original container tightly closed. Store prepared plates and the lyophilized supplements at 2-8°C away from light. Do not use the product beyond its expiry date on the label or if product shows any evidence of contamination or any sign of deterioration. © Liofilchem ® - Chromatic TM ESBL and Chromatic TM ESBL+AmpC - Rev.0 / 19.03.2014 Page 1 of 2 Instructions For Use

Transcript of © Liofilchem - ChromaticTM ESBL and ChromaticTM ESBL+AmpC...

ChromaticTM ESBLChromaticTM ESBL+AmpC

Chromogenic media for detection of ESBLs and AmpC in Enterobacteriaceae directly from clinical specimens.

ENGLISHChromaticTM ESBLChromaticTM ESBL+AmpC

Chromogenic media for detection of ESBLs and AmpC in Enterobacteriaceae directly from clinical specimens.

DESCRIPTIONChromaticTM ESBL is a chromogenic medium used for detection of Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. ESBLs are enzymes that hydrolyze most penicillins and cephalosporins, inhibited by β-lactamase inhibitors such as clavulanic acid, sulbactam and tazobactam.

ChromaticTM ESBL+AmpC is a chromogenic medium used for detection of ESBL and AmpC producers. AmpC-type enzymes hydrolyze penicillins, cephalosporins and monobactam and are poorly inhibited by the classical ESBL inhibitors, especially clavulanic acid.

ESBL-producing Enterobacteriaceae have become one of the most important causes of nosocomial community-acquired infections caused by Escherichia coli and Klebsiella pneumoniae, but also other Gram-negative bacteria. Patients infected by ESBL producers often received inadequate empirical therapy until the pathogen’s resistance is recognized. Early detection is, therefore, essential in order to limit the spread of these pathogens.Organisms producing enough AmpC β-lactamase will typically give a positive ESBL screening test but fail the confirmatory test involving increased sensitivity with clavulanic acid.

TYPICAL FORMULA (g/l)

Peptone Mix 43.2

Chromogenic Mix 1.0

Agar 15.0

Selective Mix 0.5

Final pH 7.2 ± 0.2 at 25°C

METHOD PRINCIPLEPeptones supply amino acids, nitrogen, carbon, minerals, vitamins and other nutrients which support the growth of microorganisms. Chromogenic mix allows the identification of microorganisms on the basis of the colony color and morphology. Agar is the solidifying agent.

The Selective mix in Chromatic™ ESBL inhibits the ESBL-non-producing organisms, including AmpC producers. The Selective mix in Chromatic™ ESBL+AmpC inhibits the ESBL-non-producing organisms, but allows the growth of AmpC producers.

PREPARATIONDehydrated medium Suspend 29.6 g of the powder in 500 milliliters of distilled or deionized water. Mix well. Heat

until completely dissolved. Sterilize in autoclave at 121°C for 15 minutes. Cool to 45-50°C. Aseptically, add 5 ml previously reconstituted supplement, mix well and dispense in Petri dishes.

Supplements Add 5 ml distilled or deionized water to the lyophilized supplement tube to be used. Mix well using a vortex mixer before adding to the medium being prepared.

TEST PROCEDUREInoculate the plates by streaking directly the specimen onto the agar surface. Incubate aerobically at 37°C for 18-24 hours.

INTERPRETING RESULTSAfter incubation observe the color of the colonies and interpret the results as indicated in the ID table.

ID Table.

Microorganism* Typical colony color

E. coli Pink-reddish-mauve

Klebsiella spp, Enterobacter spp, Serratia spp Green-blue

Proteus spp Brown

*On ChromaticTM ESBL medium only ESBL-producing organisms grow, while ChromaticTM ESBL+AmpC allows the growth of both ESBL and AmpC producers.

See pictures in Appendix I.

APPEARANCEDehydrated medium: fine, dry, homogeneous, free of extraneous material, beige. Lyophilized supplements: whitish button, limpid solution once reconstituted.Prepared medium: slightly opalescent, amber.

STORAGEThe powder is very hygroscopic, store the powder at 10-30°C, in a dry environment, in its original container tightly closed. Store prepared plates and the lyophilized supplements at 2-8°C away from light. Do not use the product beyond its expiry date on the label or if product shows any evidence of contamination or any sign of deterioration.

© Liofilchem® - ChromaticTM ESBL and ChromaticTM ESBL+AmpC - Rev.0 / 19.03.2014

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Instructions For Use

SHELF LIFEDehydrated medium: 4 years.Lyophilized supplements: 4 years.Ready-to-use plates: 4 months.

QUALITY CONTROLPlates are inoculated with the microbial strains indicated in the QC tables. Inoculum for productivity: 10-100 CFU/ml.Inoculum for selectivity: 104-105 CFU/ml.Incubation conditions: aerobically at 35±2°C for 18-24 h.

ChromaticTM ESBL, QC Table.ChromaticTM ESBL, QC Table.

Microorganism ESBL phenotype

AmpCphenotype Growth Specification

Escherichia coli DSM 22311 Positive Negative Good Reddish coloniesKlebsiella pneumoniae ATCC® 700603 Positive Negative Good Green-blue coloniesKlebsiella pneumoniae ATCC® BAA-1144 Negative Positive Inhibited ---Escherichia coli ATCC® 25922 Negative Negative Inhibited ---

ChromaticTM ESBL+AmpC, QC Table.ChromaticTM ESBL+AmpC, QC Table.

Microorganism ESBLphenotype

AmpCphenotype Growth Specification

Enterobacter cloacae ATCC® BAA-1143 Negative Positive Good Green-blue coloniesKlebsiella pneumoniae ATCC® 700603 Positive Negative Good Green-blue coloniesKlebsiella pneumoniae ATCC® BAA-1144 Negative Positive Good Green-blue coloniesEscherichia coli ATCC® 25922 Negative Negative Inhibited ---

WARNING AND PRECAUTIONSThe product does not contain hazardous substances in concentrations exceeding the limits set by current legislation and therefore is not classified as dangerous. It is nevertheless recommended to consult the safety data sheet for its correct use. The product is intended for In vitro diagnostic use and must be used only by properly trained operators.

DISPOSAL OF WASTEDisposal of waste must be carried out according to national and local regulations in force.

BIBLIOGRAPHY1. EUCAST guidelines for detection of resistance mechanisms and specific resistances of clinical and/or epidemiological

importance. Version 1.0, 2013.2. Podschun R, Ullman U. Klebsiella spp as Nosocomial Pathogens: Epidemiology, Taxonomy, Typing Methods, and

Pathogenicity Factors. Clinical Microbiology Reviews. 1998; 11 (4): 589–603.3. Geiss H.K. Comparison of two test kits for rapid identification of Escherichia coli by a beta-glucuronidase assay.

European Journal of Clinical Microbiology & Infections Diseases. 1990; 9 (2):151-152.

PRESENTATION Contents Ref.

ChromaticTM ESBL 90 mm ready-to-use plates 20 plates 11622

ChromaticTM ESBL+AmpC 90 mm ready-to-use plates 20 plates 11629

ChromaticTM ESBL Dehydrated medium 500 g of powder 610629

ChromaticTM ESBL Dehydrated medium 100 g of powder 620629

ChromaticTM ESBL Supplement Lyophilized supplement 10 tubes 81089

ChromaticTM ESBL+AmpC Supplement Lyophilized supplement 10 tubes 81090

TABLE OF SYMBOLS

LOT Batch code IVD In vitro Diagnostic Medical Device Manufacturer Use by Fragile, handle

with careKeep away from sunlight

REF Catalogue number

Temperature limitation

Contains sufficient for <n> tests

Caution, consult Instruction For Use

Do not reuse

LIOFILCHEM® s.r.l. Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected]

© Liofilchem® - ChromaticTM ESBL and ChromaticTM ESBL+AmpC - Rev.0 / 19.03.2014

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Instructions For Use

Istruzioni per l’uso

Chromatic™ ESBLChromatic™ ESBL+AmpC

Terreno cromogenico per la ricerca di ESBL ed AmpC in Enterobacteriaceae direttamente da campioni clinici.

ITALIANOChromatic™ ESBLChromatic™ ESBL+AmpC

Terreno cromogenico per la ricerca di ESBL ed AmpC in Enterobacteriaceae direttamente da campioni clinici.

DESCRIZIONEChromatic™ ESBL è un terreno cromogenico utilizzato per la ricerca di Enterobacteriaceae produttrici di β-lattamasi a spettro esteso. ESBL sono enzimi che idrolizzano la maggior parte delle penicilline e cefalosporine, inibiti da inibitori delle β-lattamasi come acido clavulanico, sulbactam e tazobactam.

Chromatic™ ESBL+AmpC è un terreno cromogenico utilizzato per la ricerca di produttori di ESBL ed AmpC. AmpC sono enzimi che idrolizzano penicilline, cefalosporine e monobattami e sono inibiti debolmente dai classici inibitori degli ESBL, specialmente l’acido clavulanico.

Le Enterobacteriaceae produttrici di ESBL sono diventate tra i maggiori responsabili di infezioni nosocomiali principalmente ad opera di Escherichia coli e Klebsiella pneumoniae, ma anche altri batteri Gram-negativi. Pazienti infettati da produttori di ESBL spesso ricevono terapie empiriche inadeguate finché la resistenza del patogeno non viene riconosciuta. L'identificazione precoce è, quindi, essenziale al fine di limitare la diffusione di questi patogeni.

Gli organismi che producono abbastanza β-lattamasi AmpC tipicamente risultano positivi ad uno screening per ESBL ma falliscono i test di conferma che prevedono un aumento di sensibilità con l’acido clavulanico.

FORMULA TIPICA (g/l)

Miscela di Peptoni 43.2

Miscela Cromogenica 1.0

Agar 15.0

Miscela Selettiva 0.5

pH Finale 7.2 ± 0.2 a 25°C

PRINCIPIO DEL METODOI peptoni forniscono amino acidi, azoto, carbonio, minerali, vitamine ed altri nutrienti che supportano la crescita dei microrganismi. La miscela cromogenica permette l’identificazione dei microrganismi sulla base del colore e della morfologia delle colonie. L’agar è l’agente solidificante.

La Miscela selettiva in Chromatic™ ESBL inibisce gli organismi che non producono ESBL, inclusi i produttori di AmpC. La Miscela Selettiva in Chromatic™ ESBL+AmpC inibisce gli organismi che non producono ESBL, ma permette la crescita dei produttori di AmpC.

PREPARAZIONETerreno disidratato Sospendere 29.6 g di polvere in 500 millilitri di acqua distillata o deionizzata sterile. Mescolare

bene. Riscaldare fino a completa dissoluzione. Sterilizzare in autoclave a 121°C per 15 minuti. Raffreddare il terreno a 45-50°C. In condizioni di asepsi, aggiungere 5 ml di supplemento precedentemente ricostituito, mescolare bene e dispensare in piastre Petri.

Supplementi Aggiungere 5 ml di acqua distillata o deionizzata nella provetta contenente il supplemento liofilizzato da utilizzare. Mescolare bene utilizzando un miscelatore vortex prima di aggiungere al terreno in preparazione.

PROCEDURA DEL TESTInoculare la piastra strisciando direttamente il campione clinico sulla superficie dell’agar. Incubare in atmosfera aerobica a 37°C per 18-24 ore.

INTERPRETAZIONE DEI RISULTATIDopo l’incubazione osservare il colore delle colonie ed interpretare i risultati come indicato nella tabella ID.

Tabella ID.

Microrganismo* Colore tipico delle colonie

E. coli Rosa-rossastro-malva

Klebsiella spp, Enterobacter spp, Serratia spp Verde-blu

Proteus spp Marrone

*Sul terreno Chromatic™ ESBL crescono solo organismi produttori di ESBL, mentre Chromatic™ ESBL+AmpC permette la crescita di produttori sia di ESBL che AmpC.

Consultare le figure nell’Appendice I.

ASPETTOTerreno disidratato: fine, asciutto, omogeneo, privo di materiale estraneo, beige. Supplemento liofilizzato: bottone biancastro, soluzione limpida una volta ricostituita.Terreno preparato: ambra, leggermente opalescente.

© Liofilchem® - Chromatic™ ESBL and Chromatic™ ESBL+AmpC - Rev.0 / 19.03.2014

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CONSERVAZIONELa polvere è fortemente igroscopica, conservare a 10-30°C, in ambiente asciutto, nel suo contenitore originale chiuso ermeticamente. Conservare le piastre pronte ed i supplementi liofilizzati a 2-8°C al riparo dalla luce. Non usare il prodotto dopo la sua data di scadenza indicata sull’etichetta o se il prodotto mostra segni di contaminazione o deterioramento.

DURATA Terreno disidratato: 4 anni.Supplementi liofilizzati: 4 anni.Piastre pronte all’uso: 4 mesi.

CONTROLLO DI QUALITÀLe piastre vengono inoculate con i ceppi microbici indicati nelle tabelle CQ. Inoculo per produttività: 10-100 UFC/ml.Inoculo per selettività: 104-105 UFC/ml.Condizioni di incubazione: ambiente aerobico a 35±2°C per 18-24 ore.

ChromaticTM ESBL, Tabella CQ.ChromaticTM ESBL, Tabella CQ.

Microrganismo Fenotipo ESBL

Fenotipo AmpC Crescita Specifiche

Escherichia coli DSM 22311 Positivo Negativo Buona Colonie rossastreKlebsiella pneumoniae ATCC® 700603 Positivo Negativo Buona Colonie verdi-bluKlebsiella pneumoniae ATCC® BAA-1144 Negativo Positivo Inibita ---Escherichia coli ATCC® 25922 Negativo Negativo Inibita ---

ChromaticTM ESBL+AmpC, QC Tabella CQ.ChromaticTM ESBL+AmpC, QC Tabella CQ.

Microrganismo Fenotipo ESBL

Fenotipo AmpC Crescita Specifiche

Enterobacter cloacae ATCC® BAA-1143 Negativo Positivo Buona Colonie verdi-bluKlebsiella pneumoniae ATCC® 700603 Positivo Negativo Buona Colonie verdi-bluKlebsiella pneumoniae ATCC® BAA-1144 Negativo Positivo Buona Colonie verdi-bluEscherichia coli ATCC® 25922 Negativo Negativo Inibita ---

AVVERTENZE E PRECAUZIONIIl prodotto non contiene sostanza nocive in concentrazioni superiori ai limiti fissati dall’attuale legislazione e perciò non è classificato come pericoloso. Ciononostante si raccomanda di consultare la scheda di sicurezza per il suo corretto uso. Il prodotto è da intendersi per uso diagnostico in vitro e deve essere utilizzato esclusivamente da operatori adeguatamente addestrati.

SMALTIMENTO DEI RIFIUTILo smaltimento dei rifiuti deve essere effettuato in conformità alle normative nazionali e locali in vigore.

BIBLIOGRAFIA1. EUCAST guidelines for detection of resistance mechanisms and specific resistances of clinical and/or epidemiological

importance. Version 1.0, 2013.2. Podschun R, Ullman U. Klebsiella spp as Nosocomial Pathogens: Epidemiology, Taxonomy, Typing Methods, and

Pathogenicity Factors. Clinical Microbiology Reviews. 1998; 11 (4): 589–603.3. Geiss H.K. Comparison of two test kits for rapid identification of Escherichia coli by a beta-glucuronidase assay.

European Journal of Clinical Microbiology & Infections Diseases. 1990; 9 (2):151-152.

PRESENTAZIONE Contenuto Ref.

ChromaticTM ESBL Piastre da 90 mm pronte all’uso 20 piastre 11622

ChromaticTM ESBL+AmpC Piastre da 90 mm pronte all’uso 20 piastre 11629

ChromaticTM ESBL Terreno disidratato 500 g di polvere 610629

ChromaticTM ESBL Terreno disidratato 100 g di polvere 620629

ChromaticTM ESBL Supplement Supplemento liofilizzato 10 provette 81089

ChromaticTM ESBL+AmpC Supplement Supplemento liofilizzato 10 provette 81090

TABELLA DEI SIMBOLI

LOT Codice del lotto IVD Dispositivo Medico

Diagnostico in vitroFabbricante Utilizzare entro Fragile, maneggiare

con cura

REF Numero di catalogo Limiti di temperatura Contenuto sufficiente per

<n> saggiAttenzione, Consultare le istruzioni per l’uso

Non riutilizzare

© Liofilchem® - Chromatic™ ESBL and Chromatic™ ESBL+AmpC - Rev.0 / 19.03.2014

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Istruzioni per l’uso

LIOFILCHEM® s.r.l. Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected]

ChromaticTM ESBLChromaticTM ESBL+AmpC

Chromogenic media for detection of ESBL and AmpC in Enterobacteriaceae directly from clinical specimens.

ENGLISHChromaticTM ESBLChromaticTM ESBL+AmpC

Chromogenic media for detection of ESBL and AmpC in Enterobacteriaceae directly from clinical specimens.

Appendix IChromaticTM ESBL

ChromaticTM ESBL+AmpCChromogenic media for detection of ESBL and AmpC

in Enterobacteriaceae directly from clinical specimens.

Colony appearance on ChromaticTM ESBL and ChromaticTM ESBL+AmpC plates.

© Liofilchem® - ChromaticTM ESBL and ChromaticTM ESBL+AmpC - Rev.0 / 19.03.2014

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LIOFILCHEM® s.r.l. Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected] Via Scozia zona ind.le, 64026 Roseto degli Abruzzi (Te) Italy Tel. +39 0858930745 Fax +39 0858930330 www.liofilchem.net [email protected]

Instructions For Use

E. coli DSM 22311 (ESBL+) K. pneumoniae ATCC® 700603 (ESBL+)