Microbiology Methods for ERP Review 3-2020

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B. Apparatus

Refrigerator .— capable of maintaining 2–8°C.

( a )

( b ) Incubators .— capable of maintaining 37 ± 1°C and 41.5 ± 1°C. Circulating water bath .— capable of maintaining 42–50°C. ( d ) Heating block or water bath .— capable of maintaining 85–100°C. ( c )

( e ) ( f ) (g)

Autoclave for decontamination of samples .

Homogenizer.

Measuring cylinder .— capable of measuring 2 L.

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( h ) Vessel or container .— capable of storing 2 L.

( i )

Filter laboratory blender bags .

( j ) Sterile collection sponge and swabs for sampling environmental surfaces . ( k ) Serological Pipette Bulbs (Automatic Pipette) .— For sampling and delivering of 1–10 mL.

Serological pipettes .— Aerosol resistant.

( l )

( m ) Precision pipettes .— For sampling and delivering of 100–300 µL, and 200–1,000 µL.

( n ) Micropipette tips .— Aerosol resistant.

( o ) Multi ‐ channel pipette .— Capable of delivering 100–300 µL.

( p ) Microplate washer .

( q ) Microplate reader with 450nm filter .

( r ) Calibrated Thermometers .— Capable of measuring a temperature of 37 ± 1°C, 41.5 ± 1°C, and

85–100°C.

Vortex mixer .— For swab homogenization.

( s ) ( t )

Calibrated Timer .

( u ) Marker .

C. Media and Reagents

( a ) One or Five x 96 well microplates (in breakable strip format) .— Wells are coated with

antibodies against Salmonella species. Store at 2–8°C.

( b ) Negative Control (Green Label) .— 3 mL or 10 mL in working dilution. Contains diluent with

preservative. Store at 2–8°C.

( c ) Positive Control (Red Label) .— 3 mL or 10 mL in working dilution. Contains heat ‐ killed 33 Salmonella Agona in diluent with preservative. The positive control is black in color. Store at 2–8°C. 34 ( d ) Conjugate (Orange Label) .— 11 mL or 60 mL in working dilution. Contains horseradish 35 peroxidase ‐ antibody conjugate in diluent with preservative. Store at 2–8°C. 36 ( e ) Substrate (Blue Label) .— 11 mL or 60 mL in working dilution. Contains 3,3’,5,5’ ‐ 37 Tetramethylbenzidine (TMB), hydrogen peroxide and stabilizers. Note: Solution should be colorless. 38 Store at 2–8°C. 39 ( f ) Stop Solution (Silver Label) .— 11 mL or 60 mL in working dilution and contains 0.2M sulfuric 40

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