GB 4789.30-2010 English PDF

GB 4789.30-2010: National food safety standard -- Food microbiological examination: Listeria monocytogenes
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GB 4789.30-2010
GB
National Standard of the People’s Republic of China
National food safety standard
Food microbiological examination.
Listeria monocytogenes
ISSUED ON. MARCH 26, 2010
IMPLEMENTED ON. JUNE 01, 2010
Issued by. Ministry of Health of the People’s Republic of China
Table of Contents
Foreword ... 3 
1 Scope ... 4 
2 Apparatuses and materials ... 4 
3 Culture medium and reagents ... 5 
4 Inspection procedure ... 5 
5 Operation steps ... 6 
6 Results and report ... 8 
Appendix A ... 9 
Foreword 
This standard will replace GB/T 4789.30-2008 Microbiological examination of food
hygiene - Examination of listeria monocytogenes.
Compared with GB/T 4789.30-2008, the main changes of this standard are as follows.
- Modify this standard’s Chinese and English names;
- Delete “the second method - Automatic enzyme-linked fluorescence
immunoassay analyzer screening method”;
- Delete “the third method - Automatic screening method of pathogen detection
system”.
This standard’s appendix A is normative.
The previous versions replaced by this standard are.
- GB 4789.30-1994, GB/T 4789.30-2003, GB/T 4789.30-2008.
National food safety standard
Food microbiological examination.
Listeria monocytogenes
1 Scope
This standard specifies the test methods of Listeria monocytogenes in foods.
This standard applies to test of Listeria monocytogenes in foods.
2 Apparatuses and materials   
In addition to the conventional sterilization and cultivation apparatuses, other
apparatuses and materials are as follows.
2.1 Refrigerator. 2°C~5°C.
2.2 Thermostatic incubator. 30°C±1°C, 36°C±1°C.
2.3 Homogenizer.
2.4 Microscope. 10x~100x.
2.5 Electronic balance. Sensitivity is 0.1g.
2.6 Erlenmeyer flasks. Capacity 100mL and 500mL.
2.7 Sterile pipette. 1mL (with 0.01mL scale), 10mL (with 0.1mL scale).
2.8 Sterile petri dish. Diameter is 90mm.
2.9 Sterile test tube. 16mmx160mm.
2.10 Centrifuge tube. 30mmx100mm.
2.11 Sterile syringe. 1mL.
2.12 Staphylococcus aureus (ATCC25923).
2.13 Rhodococcus equi.
2.14 Mus musculus albus. 16g - 18g.
2.15 Automated microbial and biochemical identification system.
3 Culture medium and reagents   
3.1 Tryptone soya broth containing 0.6% yeast extract (TSB-YE). see appendix A.1.
3.2 Trypticase soy agar containing 0.6% yeast extract (TSA-YE). see appendix A.2.
3.3 Listeria enrichment broth LB (LB1, LB2). see appendix A.3.
3.4 1% acriflavine HCL solution. see appendix A.3.2.1.
3.5 1% of naladixic acid solution. see appendix A.3.2.1.
3.6 PALCAM agar. see appendix A.4.
3.7 Gram dye. see appendix A.5.
3.8 SIM motility culture medium. see appendix A.6.
3.9 Buffer glucose peptone water [for Methyl Red (MR) and V-P tests]. see appendix
A.7.
3.10 5%~8% sheep blood agar. see appendix A.8.
3.11 Sugar fermentation tube. see appendix A.9.
3.12 Catalase test. see appendix A.10.
3.13 Listeria chromogenic medium.
3.14 Biochemical identification kit.
4 Inspection procedure 
The inspection procedure of Listeria monocytogenes is shown in Figure 1.
A.9.2 Preparation method
A.9.2.1 Allocate glucose fermentation tubes according to above compositions. Add
glucose according to proportion 0.5%. Pack into the small test tubes which have
inverted tubules. Adjust pH to 7.4. Autoclave it for 15min at 115°C. Preserve for spare
use.
A.9.2.2 Allocate other glucose fermentation tubes according to above compositions.
Pack every bottle for 100mL. Autoclave it for 15min at 115°C. Allocate all types of
carbohydrate to make 10% solution. Autoclave it simultaneously. Add 5mL of sugar
solution to 100mL of culture medium. Use sterile technique to pack into small tubes.
A.9.3 Test method
Use appropriate pure cultures to inoculate in sugar fermentation tube. Cultivate it for
24h~48h at 36°C±1°C. Observe the result. Blue represents negative. Yellow
represents positive.
A.10 Catalase test
A.10.1 Reagents
3% Hydrogen peroxide solution. Prepare it when use.
A.10.2 Test method
Use small glass rod or disposable inoculation needle to pick single colony. Place it in
clean test tube. Add 2mL of 3% Hydrogen peroxide solution. Observe the result.
A.10.3 Results
If it produces bubbles within half minute, the result is positive. If not, the result is
negative.
GB 4789.30-2010
GB
National Standard of the People’s Republic of China
National food safety standard
Food microbiological examination.
Listeria monocytogenes
ISSUED ON. MARCH 26, 2010
IMPLEMENTED ON. JUNE 01, 2010
Issued by. Ministry of Health of the People’s Republic of China
Table of Contents
Foreword ... 3 
1 Scope ... 4 
2 Apparatuses and materials ... 4 
3 Culture medium and reagents ... 5 
4 Inspection procedure ... 5 
5 Operation steps ... 6 
6 Results and report ... 8 
Appendix A ... 9 
Foreword 
This standard will replace GB/T 4789.30-2008 Microbiological examination of food
hygiene - Examination of listeria monocytogenes.
Compared with GB/T 4789.30-2008, the main changes of this standard are as follows.
- Modify this standard’s Chinese and English names;
- Delete “the second method - Automatic enzyme-linked fluorescence
immunoassay analyzer screening method”;
- Delete “the third method - Automatic screening method of pathogen detection
system”.
This standard’s appendix A is normative.
The previous versions replaced by this standard are.
- GB 4789.30-1994, GB/T 4789.30-2003, GB/T 4789.30-2008.
National food safety standard
Food microbiological examination.
Listeria monocytogenes
1 Scope
This standard specifies the test methods of Listeria monocytogenes in foods.
This standard applies to test of Listeria monocytogenes in foods.
2 Apparatuses and materials   
In addition to the conventional sterilization and cultivation apparatuses, other
apparatuses and materials are as follows.
2.1 Refrigerator. 2°C~5°C.
2.2 Thermostatic incubator. 30°C±1°C, 36°C±1°C.
2.3 Homogenizer.
2.4 Microscope. 10x~100x.
2.5 Electronic balance. Sensitivity is 0.1g.
2.6 Erlenmeyer flasks. Capacity 100mL and 500mL.
2.7 Sterile pipette. 1mL (with 0.01mL scale), 10mL (with 0.1mL scale).
2.8 Sterile petri dish. Diameter is 90mm.
2.9 Sterile test tube. 16mmx160mm.
2.10 Centrifuge tube. 30mmx100mm.
2.11 Sterile syringe. 1mL.
2.12 Staphylococcus aureus (ATCC25923).
2.13 Rhodococcus equi.
2.14 Mus musculus albus. 16g - 18g.
2.15 Automated microbial and biochemical identification system.
3 Culture medium and reagents   
3.1 Tryptone soya broth containing 0.6% yeast extract (TSB-YE). see appendix A.1.
3.2 Trypticase soy agar containing 0.6% yeast extract (TSA-YE). see appendix A.2.
3.3 Listeria enrichment broth LB (LB1, LB2). see appendix A.3.
3.4 1% acriflavine HCL solution. see appendix A.3.2.1.
3.5 1% of naladixic acid solution. see appendix A.3.2.1.
3.6 PALCAM agar. see appendix A.4.
3.7 Gram dye. see appendix A.5.
3.8 SIM motility culture medium. see appendix A.6.
3.9 Buffer glucose peptone water [for Methyl Red (MR) and V-P tests]. see appendix
A.7.
3.10 5%~8% sheep blood agar. see appendix A.8.
3.11 Sugar fermentation tube. see appendix A.9.
3.12 Catalase test. see appendix A.10.
3.13 Listeria chromogenic medium.
3.14 Biochemical identification kit.
4 Inspection procedure 
The inspection procedure of Listeria monocytogenes is shown in Figure 1.
A.9.2 Preparation method
A.9.2.1 Allocate glucose fermentation tubes according to above compositions. Add
glucose according to proportion 0.5%. Pack into the small test tubes which have
inverted tubules. Adjust pH to 7.4. Autoclave it for 15min at 115°C. Preserve for spare
use.
A.9.2.2 Allocate other glucose fermentation tubes according to above compositions.
Pack every bottle for 100mL. Autoclave it for 15min at 115°C. Allocate all types of
carbohydrate to make 10% solution. Autoclave it simultaneously. Add 5mL of sugar
solution to 100mL of culture medium. Use sterile technique to pack into small tubes.
A.9.3 Test method
Use appropriate pure cultures to inoculate in sugar fermentation tube. Cultivate it for
24h~48h at 36°C±1°C. Observe the result. Blue represents negative. Yellow
represents positive.
A.10 Catalase test
A.10.1 Reagents
3% Hydrogen peroxide solution. Prepare it when use.
A.10.2 Test method
Use small glass rod or disposable inoculation needle to pick single colony. Place it in
clean test tube. Add 2mL of 3% Hydrogen peroxide solution. Observe the result.
A.10.3 Results
If it produces bubbles within half minute, the result is positive. If not, the result is
negative.