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GB 4789.30-2016 English PDF (GB 4789.30-2010, GB/T 4789.30-2008)

GB 4789.30-2016_English: PDF (GB4789.30-2016)
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GB 4789.30-2016English115 Add to Cart 0--9 seconds. Auto-delivery National Food Safety Standard -- Food Microbiological Examination -- Examination of Listeria Monocytogenes Valid GB 4789.30-2016
GB 4789.30-2010English70 Add to Cart 0--9 seconds. Auto-delivery National food safety standard -- Food microbiological examination: Listeria monocytogenes Obsolete GB 4789.30-2010
GB/T 4789.30-2008English679 Add to Cart 5 days [Need to translate] Microbiological examination of food hygiene -- Examination of listeria monocytogenes Obsolete GB/T 4789.30-2008
GB/T 4789.30-2003English359 Add to Cart 3 days [Need to translate] Microbiological examination of food hygiene -- Examination of listeria monocytogenes Obsolete GB/T 4789.30-2003
GB 4789.30-1994EnglishRFQ ASK 3 days [Need to translate] Microbiological examination of food hygiene. Examination of Listeria moncytogenes Obsolete GB 4789.30-1994


BASIC DATA
Standard ID GB 4789.30-2016 (GB4789.30-2016)
Description (Translated English) National Food Safety Standard -- Food Microbiological Examination -- Examination of Listeria Monocytogenes
Sector / Industry National Standard
Classification of Chinese Standard X09
Word Count Estimation 16,187
Date of Issue 2016-12-23
Date of Implementation 2017-06-23
Older Standard (superseded by this standard) GB 4789.30-2010
Regulation (derived from) National Health and Family Planning Commission Notice No.17 of 2016

BASIC DATA
Standard ID GB 4789.30-2010 (GB4789.30-2010)
Description (Translated English) National food safety standard - Food microbiological examination: Listeria monocytogenes
Sector / Industry National Standard
Classification of Chinese Standard C53
Classification of International Standard 07.100.30
Word Count Estimation 10,153
Date of Issue 2010-03-26
Date of Implementation 2010-06-01
Older Standard (superseded by this standard) GB/T 4789.30-2008
Regulation (derived from) Circular of the Ministry of Health (2010)7
Issuing agency(ies) National Health and Family Planning Commission of People's Republic of China
Summary This Chinese standard specifies the food Listeria monocytogenes bacteria (Listeria monocytogenes) test. This standard applies to foods Listeria monocytogenes bacteria test.

BASIC DATA
Standard ID GB/T 4789.30-2008 (GB/T4789.30-2008)
Description (Translated English) Microbiological examination of food hygiene. Examination of listeria monocytogenes
Sector / Industry National Standard (Recommended)
Classification of Chinese Standard C53
Classification of International Standard 07.100.30
Word Count Estimation 17,178
Date of Issue 2008-11-21
Date of Implementation 2009-03-01
Older Standard (superseded by this standard) GB/T 4789.30-2003
Drafting Organization China Disease Prevention and Control Center of Nutrition and Food Safety
Administrative Organization Ministry of Health
Regulation (derived from) National Standard Approval Announcement 2008 No.22 (Total No.135); Health-Communication (2010) 7; National Standard Approval Announcement 2010 No.5 (Total No.160)
Proposing organization People's Republic of China Ministry of Health
Issuing agency(ies) Ministry of Health of People's Republic of China; Standardization Administration of China
Summary This standard specifies the foods monocytogenes Listeria (Listeria monocytogenes) test methods. This standard applies to food and foodborne disease samples monocytogenes Listeria test.


GB 4789.30-2016 GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA National food safety standard - Food microbiological examination - Examination of Listeria monocytogenes ISSUED ON. DECEMBER 23, 2016 IMPLEMENTED ON. JUNE 23, 2017 Issued by. National Health and Family Planning Commission; China Food and Drug Administration. Table of Contents Foreword ... 3  1 Scope ... 4  2 Equipment and materials ... 4  3 Media and reagents ... 5  4 Testing procedures ... 6  5 Operation procedures ... 6  6 Test procedures ... 9  7 Operation procedures ... 9  8 Result count ... 11  9 Results report ... 12  10 Testing procedures ... 12  11 Operation procedures... 13  12 Results and reports ... 14  Appendix A Medium and reagents ... 15  Appendix B Listeria monocytogenes most probable number (MPN) search table ... 23  National food safety standard - Food microbiological examination - Examination of Listeria monocytogenes 1 Scope This standard specifies the test method for Listeria monocytogenes in food. The first method of this standard is applicable to the qualitative test of Listeria monocytogenes in food; the second method is applicable to the counting of Listeria monocytogenes in foods with higher Listeria monocytogenes; the third method is applicable to the count of Listeria monocytogenes in foods with low Listeria monocytogenes (< 100 CFU/g) and high bacterial content, especially milk, water and food containing particulate matter which interferes with the counting of colonies. 2 Equipment and materials In addition to routine sterilization and culture equipment in microbiology laboratories, other equipment and materials are as follows. 2.1 Refrigerator. 2 °C ~ 5 °C. 2.2 Constant temperature incubator. 30 °C ± 1 °C, 36 °C ± 1 °C. 2.3 Homogenizer. 2.4 Microscope. 10 x ~ 100 x. 2.5 Electronic balance. the sensitivity is 0.1 g. 2.6 Conical flask. 100 mL, 500 mL. 2.7 Sterile pipette. 1 mL (with 0.01 mL scale), 10 mL (with 0.1 mL scale) or micropipette and tip. 2.8 Sterile plate. 90 mm in diameter. 2.9 Sterile test tube. 16 mm × 160 mm. 2.10 Centrifuge tube. 30 mm × 100 mm. 2.11 Sterile syringe. 1 mL. HOMOGENIZE it at 8000 r/min ~ 10000 r/min for 1 min ~ 2 min. CULTURE it at 30 °C ± 1 °C for 24 h ± 2 h, TAKE 0.1 mL, TRANSFER it to 10 mL of LB2 monocytogenes solution, CULTURE it at 30 °C ± 1 °C for 24 h ± 2 h. 5.2 Separation TAKE the LB2 secondary monocytogenes solution, INOCULATE it onto the Listeria chromogenic plate and the PALCAM agar plate in a line shape, CULTURE it at 36 °C ± 1 °C for 24 h ~ 48 h, OBSERVE the colonies growing on each plate. Typical colonies are small round gray-green colonies on PALCAM agar plates with brown-black hydrolyzed circles around them, some colonies have black depressions; colony characteristics on Listeria color- developing plates are determined by reference to product specifications. 5.3 Initial screening TAKE to five typical or suspect colonies from selective agar plates, respective INOCULATE them into the xylose and rhamnose fermentation tubes, CULTURE it at 36 °C ± 1 °C for 24 h ± 2 h, meanwhile MARK a line on the TSA-YE plate, CULTURE it at 36 ° C ± 1 ° C for 18 h ~ 24 h, then SELECT the pure culture of xylose-negative and rhamnose-positive for further identification. 5.4 Identification (or selection of biochemical identification kits or fully automated microbial identification systems, etc.) 5.4.1 Staining microscopy. Listeria monocytogenes is a Gram-positive Brevibacterium, the size is (0.4 μm ~ 0.5 μm) × (0.5 μm ~ 2.0 μm); the bacterial suspension is made with physiological saline, which is observed in oil mirror or under the phase contrast microscope, the bacteria shows slight rotation or rolling-like motion. 5.4.2 Dynamic test. PICK a single suspicious colony to puncture semi-solid or SIM dynamic medium in pure culture, CULTURE it at 25 °C ~ 30 °C for 48 h. Listeria is motivated and has an umbrella shape growth above semi-solid or SIM medium. If the umbrella growth is not obvious, it can continue to culture it for 5 d, then observe the results. 5.4.3 Biochemical identification. PICK the single suspicious colony in pure culture to perform catalase test. Colonies with positive catalase reaction is continued to be subject to sugar fermentation test and MR-VP test. The main biochemical characteristics of Listeria monocytogenes are as shown in Table 1. 5.4.4 Hemolysis test. DIVIDE the bottom surface of fresh sheep blood agar plate into 20 ~ 25 small cells, PICK a single suspicious colony of pure culture, INOCULATE it onto the blood plate by puncture, one colony per cell, and INOCULATE the positive control bacteria by puncture (Listeria monocytogenes, Listeria ivanovii and Listeria seeligeri) and negative control bacteria (Listeria without additives, HOMOGENIZE it continuously for 1 min ~ 2 min on a tapping homogenizer, or HOMOGENIZE it for 1 min ~ 2 min at 8000 r/min ~ 10000 r/min. SHAKE and MIX the liquid sample uniformly, to make it into 1.10 sample homogenate. 7.1.2 USE a 1 mL sterile pipette or micropipette to pipette 1 mL of 1.10 sample homogenate, along the tube wall, INJECT it slowly into a sterile test tube which contains 9 mL of buffered peptone or LB broth without additives (note avoiding the pipette or tip end from touching the dilution solution surface), SHAKE the test tube or USE another 1 mL sterile pipette to repeatedly mix it uniformly, to prepare it into 1.100 sample homogenate. 7.1.3 PREPARE 10 times serial dilution sample homogenate in accordance with the procedure of 7.1.2. For each incremental dilution, replace one 1 mL sterile pipette or tip. 7.2 Inoculation of samples In accordance with the estimation of the contamination status of the sample, SELECT 2 ~ 3 sample homogenate of appropriate continuous dilution (the liquid sample may include the original solution), respectively TAKE 1 mL of each dilution of the sample homogenate, at the inoculation amount of 0.3 mL, 0.3 mL, 0.4 mL, respectively ADD it onto three Listeria chromogenic plates, USE the sterile L bar to cover it over the entire plate, PAY attention to avoiding touching the plate edge. Before use, if there is water drops on the surface of the agar plate, it can be dried in an incubator at 25 °C ~ 50 °C until the water drops on the surface of the plate disappear. 7.3 Culture 7.3.1 Under normal circumstances, after coating, ALLOW the plate to stand for 10 min. If the sample solution is not easily absorbed, it may place the plate in an incubator at 36 °C ± 1 °C for 1 h; after the sample homogenate is absorbed, INVERT the plate, PLACE it into the incubator upside down, to culture it at 36 °C ± 1 °C for 24 h ~ 48 h. 7.4 Typical colony count and confirmation 7.4.1 Colony characteristics of Listeria monocytogenes on Listeria chromogenic plates are subject to product description. 7.4.2 SELECT a plate which has typical Listeria monocytogenes colonies, and the total number of colonies in the same dilution of 3 plates between 15 CFU and 150 CFU, COUNT the number of typical colonies. In case. a) The plate colony count of only one dilution is between 15 CFU ~ 150 CFU and there is typical colony, it shall count the typical colonies on the plate Appendix A Medium and reagents A.1 Tryptic soy broth (TSB-YE) containing 0.6% yeast extract A.1.1 Ingredients Tryptone 17.0 g Polyad tryptone 3.0 g Yeast extract 6.0 g Sodium chloride 5.0 g Dipotassium hydrogen phosphate 2.5 g Glucose 2.5 g Distilled water 1000 mL A.1.2 Preparation method HEAT and STIR the above components to dissolve it, ADJUST the pH to 7.2 ± 0.2, CONTAIN it separately, AUTOCLAVE it at 121 °C for 15 min, PREPARE for use. A.2 Tryptic soy agar (TSA-YE) containing 0.6% yeast extract A.2.1 Ingredients Tryptone 17.0 g Polyad tryptone 3.0 g Yeast extract 6.0 g Sodium chloride 5.0 g Dipotassium hydrogen phosphate 2.5 g Glucose 2.5 g Agar 15.0 g Distilled water 1000 mL Glucose 0.5 g Esculin 0.8 g Ammonium ferric citrate 0.5 g Mannitol 10.0 g Phenol red 0.1 g Lithium chloride 15.0 g Casein trypsin digest 10.0 g Heart trypsin digest 3.0 g Corn Starch 1.0 g Meat and stomach enzyme digest 5.0 g Sodium chloride 5.0 g Agar 15.0 g Distilled water 1000 mL A.4.2 Preparation method HEAT and STIR the above components to dissolve it, ADJUST the pH to 7.2 ± 0.2, CONTAIN it separately, AUTOCLAVE it at 121 °C for 15 min, PREPARE for use. A.4.2.1 PALCAM selective additives Polymyxin B 5.0 mg Acriflavine HCl 2.5 mg Ceftazidime 10.0 mg Sterile distilled water 500 mL A.4.2.2 Preparation method DISSOLVE the PALCAM base medium, COOL it to 50 °C, ADD 2 mL of PALCAM selective additive, MIX it uniformly, POUR it into a sterile plate, PREPARE for use. A.5 Gram staining solution After dissolving it, ADJUST the pH to 7.0 ± 0.2, CONTAIN it in different test tubes, 1 mL per tube, AUTOCLAVE it at 121 ° C for 15 min, PREPARE for use. A.7.3 Methyl red (MR) test A.7.3.1 Methyl red reagent A.7.3.1.1 Ingredients Methyl red 10 mg 95% ethanol 30 mL Distilled water 20 mL A.7.3.1.2 Preparation method DISSOLVE 10 mg of methyl red in 30 mL of 95% ethanol, then ADD 20 mL of distilled water. A.7.3.1.3 Test method TAKE an appropriate amount of agar culture, INOCULATE it into buffered glucose protein water, CULTURE it at 36 °C ± 1 °C for 2 d ~ 5 d. ADD one drop of methyl red reagent, immediately OBSERVE it. Bright red is positive and yellow is negative. A.7.4 V-P test A.7.4.1 6% α-naphthol-ethanol solution Ingredients and preparation method. Take 6.0 g of α-naphthol, ADD absolute ethanol to dissolve it, MAKE its volume reach to 100 mL. A.7.4.2 40% potassium hydroxide solution Ingredients and preparation method. TAKE 40 g of potassium hydroxide, ADD distilled water to dissolve it, MAKE its volume reach to 100 mL. A.7.4.3 Test method TAKE an appropriate amount of agar culture, INOCULATE it into buffered glucose protein water, CULTURE it at 36 °C ± 1 °C for 2 d ~ 4 d. ADD 0.5 mL of 6% α-naphthol-ethanol solution and 0.2 mL of a 40% potassium hydroxide solution, SHAKE the test tube completely, OBSERVE the results. If red color appears immediately or after several minutes, it is positive reaction; if it is negative reaction, it shall be cultured continuously at 36 °C ± 1 °C for 1 h and observed. meanwhile AUTOCLAVE it. ADD 5 mL of the sugar solution into 100 mL of the medium, aseptically DISPENSE it into a small tube containing the inverted tube. Or PREPARE other sugar fermentation tubes in accordance with the preparation method of A.9.2.1 glucose fermentation tube. A.9.3 Test methods TAKE appropriate amount of pure culture, INOCULATE it into the sugar fermentation tube, CULTURE it at 36 °C ± 1 °C for 24 h ~ 48 h. OBSERVE the results, the blue color indicates negative and yellow color indicates positive. A.10 Catalase test A.10.1 Reagent 3% hydrogen peroxide solution. PREPARE it at the time of use. A.10.2 Test methods USE a thin glass rod or a disposable inoculation needle to pick a single colony, PLACE it into a clean glass plate, ADD 2 drops of 3% hydrogen peroxide solution, OBSERVE the result. A.10.3 Results Those which have bubbles in half a minute are positive, and those which do not have bubbles are negative. A.11 Buffered protein water (BPW) A.11.1 Ingredients Peptone 10.0 g Sodium chloride 5.0 g Disodium hydrogen phosphate (Na2HPO4 • 12H2O) 9.0 g Potassium dihydrogen phosphate 1.5 g Distilled water 1000 mL A.11.2 Preparation method HEAT and STIR to dissolve it, ADJUST the pH to 7.2 ± 0.2, AUTOCLAVE it at 121 °C for 15 min. ......


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/T 4789.30-2008 Microbiological examination of food hygiene.Examination of listeria monocytogenes ICS 07.100.30 C53 National Standards of People's Republic of China Replacing GB/T 4789.30-2003 Microbiological examination of food hygiene Listeria monocytogenes bacteria test Posted 2008-11-21 2009-03-01 implementation People's Republic of China Ministry of Health Standardization Administration of China released Foreword The first law revision of this standard used by the US FDA "bacterial test Handbook" (Bacteriologicalanalyticalmanual, Chapter 10,2002); second method is equivalent to using "monocytogenes Listeria test methods" international analyst Institute (AOAC-RIPer- The standard method first main difference compared with the US FDA/BAM as follows. --- LB broth enrichment medium to replace BLEB broth; --- Increased screening step; --- Culture temperature to 36 ℃ ± 1 ℃ instead of 35 ℃. This standard replaces GB/T 4789.30-2003 "Microbiological examination of food hygiene monocytogenes Listeria test." This standard compared with GB/T 4789.30-2003 The main changes are as follows. --- Remove the normative references. --- Amend the law first. Including selective isolation medium to PALCAM instead of MMA, an increase of CHROMagar --- Adds a second method, enzyme-linked fluorescence immunoassay analyzer automatic screening method. --- Adds a third method, automated pathogen detection screening system. Appendix A of this standard is a normative appendix. This standard is proposed and administered by the People's Republic of China Ministry of Health. This standard by the People's Republic of China Ministry of Health is responsible for interpretation. This standard is drafted by. China Center for Disease Control Nutrition and Food Safety. Participated in the drafting of this standard. Disease Prevention and Control Center of Fujian Province, Shaanxi Provincial Disease Prevention and Control Center, Hubei Provincial Disease Prevention and Control center. The main drafters of this standard. Xiumei, Yang Yang, Fletch, Guo Yunchang, Tian Jing, SOUTH - Ma Yi. The standard version of the calendar year instead of the standard release of case. --- GB/T 4789.30-1994, GB/T 4789.30-2003. Microbiological examination of food hygiene Listeria monocytogenes bacteria test 1 Scope This standard applies to food and food-borne illness in the sample monocytogenes Listeria test. 2 Equipment and Materials In addition to conventional microbiological laboratory culture and sterile equipment, other equipment and materials as follows. 2.1 Refrigerator. 2 ℃ ~ 5 ℃. 2.2 incubator. 30 ℃ ± 1 ℃, 36 ℃ ± 1 ℃. 2.3 homogenizer. 2.4 Microscope. 10 × ~ 100 ×. 2.5 Electronic balance. a sense of the amount of 0.1g. 2.6 conical flask. 100mL, 500mL. 2.7 sterile pipette. 1mL (with 0.01mL scale), 10mL (with 0.1mL scale). 2.8 sterile petri dish. diameter of 90mm. 2.9 sterile tube. 16mm × 160mm. 2.10 tube. 30mm × 100mm. 2.11 sterile syringes. 1mL. 2.12 Staphylococcus aureus (ATCC25923). 2.14 mice. 16g ~ 18g. 2.15 automated microbial identification system (VITEK) 1). 2.16 ELFA automatic immunoassay analyzer (miniVIDAS or VIDAS) 1). 2.17 automated pathogen detection system (BAX system, including BAX System Q7) 2). 3 media and reagents 3.1 containing 0.6% yeast extract of trypticase soy broth (TSB-YE). See Section A. Chapter 1. 1) Product name supplied by the French company bioMérieux products. This information is given for the convenience of users of this standard does not mean that the product Recognition. If other equivalent product has the same effect, you can use these equivalent products. 2) provided by the trade name of DuPont products. This information is given for the convenience of users of this standard does not imply endorsement of the product. If other equivalent product has the same effect, you can use these equivalent products. 3.2 containing 0.6% yeast extract of trypticase soy agar (TSA-YE). See Section A. 2. 3.3 Lee enrichment broth LB (LB1, LB2). See Section A. 3. 3.4 1% hydrochloric acid acridine yellow (acriflavineHCl) Solution. See A. 3.2.1. 3.5 1% nalidixic acid sodium salt (naladixicacid) Solution. See A. 3.2.1. 3.6 PALCAM Agar. see Section A. 4. 3.7 Gram stain solution. See Section A. 5. ......

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