食品中副溶血性弧菌检验能力验证样品的研制及其应用
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摘要
目的研制食品中副溶血性弧菌检验能力验证样品,并应用于副溶血性弧菌能力验证试验中。方法能力验证样品包括0～3瓶阳性样品和0～3瓶阴性样品。阴性样品仅含有背景细菌,阳性样本在背景菌的基础上添加有副溶血性弧菌。为防止数据串通,编制1～180的随机数字表,其中90个随机数字作为阳性样品编码,另外90个随机数字则用为阴性样品编码。随机抽取冻干质控阳性样品、阴性样品各20瓶,参照本次能力验证推荐的GB4789.7-2013对样品进行均匀性检验,阳性样品均需要检出副溶血性弧菌,而阴性对照样品应不得检出副溶血性弧菌。将副溶血性弧菌能力验证样品分别在?30、4℃储藏30d监测其储藏稳定性,同时在25和37℃下储藏7 d监测其运输稳定性。结果副溶血性弧菌的质控样在均匀性、储藏稳定性和运输稳定性均能满足质控样使用的要求。在39家实验室反馈结果中,36家结果评定为满意,满意率为92.3%。结论食品中副溶血性弧菌能力验证样品可以满足此次能力验证的需求,国内实验室大部分能满足考核要求,仍有部分实验室需要提高检验检测能力。
Objective To develop a verification sample for the test ability of Vibrio parahaemolyticus in food,and to apply it to the viability test of Vibrio parahaemolyticus. Methods The proficiency testing samples included 0-3 positive samples and 0-3 negative samples. The negative samples contain background bacteria only, and the positive samples were added with Vibrio parahaemolyticus on the basis of background bacteria. In order to prevent data collusion, a random number table of 1-180 was prepared, in which 90 random numbers were coded as positive samples and 90 random numbers were coded as negative samples. Totally 20 bottles of random freeze-dried positive samples and 20 bottles of random freeze-dried negative samples were randomly selected and homogeneity test was conducted on the samples with reference to the recommended standard of GB 4789.7-2013. The positive samples were detected Vibrio parahaemolyticus, and the negative control samples should be not detected Vibrio parahaemolyticus. Then the Vibrio parahaemolyticus samples were stored respectively at -30 ℃ and 4 ℃ for 30 d to evaluate the storage stability. Meanwhile the Vibrio parahaemolyticus samples were stored respectively at 25 ℃ and 37 ℃ for 7 d to evaluate the transport stability. Results The quality control samples of Vibrio parahaemolyticus could meet the requirements of using quality control samples in uniformity, storage stability and transportation stability. Among the 39 laboratory feedback results, 36 were assessed as satisfactory, with a satisfaction rate of 92.3%. Conclusion The verification sample of Vibrio parahaemolyticus in food can meet the requirements of this capability verification. Most of the domestic laboratories can meet the assessment requirements, and some laboratories still need to improve the inspection and testing capabilities.
Objective To develop a verification sample for the test ability of Vibrio parahaemolyticus in food,and to apply it to the viability test of Vibrio parahaemolyticus. Methods The proficiency testing samples included 0-3 positive samples and 0-3 negative samples. The negative samples contain background bacteria only, and the positive samples were added with Vibrio parahaemolyticus on the basis of background bacteria. In order to prevent data collusion, a random number table of 1-180 was prepared, in which 90 random numbers were coded as positive samples and 90 random numbers were coded as negative samples. Totally 20 bottles of random freeze-dried positive samples and 20 bottles of random freeze-dried negative samples were randomly selected and homogeneity test was conducted on the samples with reference to the recommended standard of GB 4789.7-2013. The positive samples were detected Vibrio parahaemolyticus, and the negative control samples should be not detected Vibrio parahaemolyticus. Then the Vibrio parahaemolyticus samples were stored respectively at -30 ℃ and 4 ℃ for 30 d to evaluate the storage stability. Meanwhile the Vibrio parahaemolyticus samples were stored respectively at 25 ℃ and 37 ℃ for 7 d to evaluate the transport stability. Results The quality control samples of Vibrio parahaemolyticus could meet the requirements of using quality control samples in uniformity, storage stability and transportation stability. Among the 39 laboratory feedback results, 36 were assessed as satisfactory, with a satisfaction rate of 92.3%. Conclusion The verification sample of Vibrio parahaemolyticus in food can meet the requirements of this capability verification. Most of the domestic laboratories can meet the assessment requirements, and some laboratories still need to improve the inspection and testing capabilities.
引文
[1]Pal D,Das N.Isolation,identification and molecular characterization of Vibrio parahaemolyticus from fish samples in Kolkata[J].Eur Rev Med Pharmacol Sci,2010,14(6):545-549.
[2]Kam KM,Luey CK,Parsons MB,et al.Evaluation and validation of a PulseNet standardized pulsed-field gel electrophoresis protocol for subtyping Vibrio parahaemolyticus:An international multicenter collaborative study[J].J Clin Microbiol,2008,46(8):2766.
[3]姜大栋,李琦.一起副溶血弧菌感染食物中毒病原学检测[J].中国现代药物应用,2018,12(7):220-222.Jiang DD,Li Q.A pathogenic test of food poisoning caused by Vibrio parahaemolyticus infection[J].Chin J Mod Drug Appl,2018,12(7):220-222.
[4]施怡茹,陈洪友,卢晓芸.一起多血清型副溶血性弧菌聚集性腹泻事件的病原分析[J].检验医学与临床,2018,12(15):3662-3666.Shi YR,Chen HY,Lu XY.Pathogen analysis of a multi-serotype Vibrio parahaemolyticus aggregation diarrhea event[J].Lab Med Clin,2018,12(15):3662-3666.
[5]Croci L.Comparison of different biochemical and molecular methods for the identification of Vibrio parahaemolyticus[J].J Appl Microbiol,2010,102(1):229-237.
[6]刘秋爽.副溶血弧菌引起食物中毒调查[J].医疗装备,2018,31(1):71-72.Liu QS.Investigation of food poisoning caused by Vibrio parahaemolyticus[J].Chin J Med Dev,2018,31(1):71-72.
[7]Laskowskiarce MA,Orth K.Acanthamoeba castellanii promotes the survival of Vibrio parahaemolyticus[J].Appl Environ Microbiol,2008,74(23):7183-7188.
[8]张万标,焦辛妮,黄坚,等.2013-2015年北海市致泻性弧菌监测情况分析[J].实用预防医学,2017,24(7):865-867.Zhang WB,Jiao XN,Huang J,et al.Analysis on the monitoring of pathogenic Vibrio in beihai from 2013 to 2015[J].Pract Prev Med,2017,24(7):865-867.
[9]季霄雷,许海燕,苏婧.82株副溶血性弧菌毒力基因及耐药性分析[J].中华微生物学和免疫学杂志,2018,38(1):37-40.Ji XL,Xu HY,Su J.Analysis of virulence genes and drug resistance in 82Vibrio parahaemolyticus strains[J].Chin J Microbiol Immunol,2018,38(1):37-40.
[10]冼钰茵,余翀,阮荣勇.广州市售水产品副溶血弧菌和溶藻弧菌的耐药性评估[J].安徽农业科学,2017,45(28):74-77.Xian YY,Yu C,Ruan RY.Drug resistance of Vibrio parahemolytics and Vibrio alginolyticus in aquatic products from guangzhou retail markets[J].J Anhui Agric Sci,2017,45(28):74-77.
[11]宫春波,王朝霞,伍海燕,等.烟台海域鲜活海产品副溶血弧菌的污染概况及致病风险评价[J].青岛农业大学学报,2017,34(3):216-221.Gong CB,Wang ZX,Wu HY,et al.Pollution situation and risk assessment of Vibrio parahaemolyticus in fresh seafood in yantai sea area[J].JQingdao Agric Univ(Nat Sci),2017,34(3):216-221.
[12]徐费霞,赵良炜,吴艳辉,等.天津地区副溶血弧菌分离株血清型、部分毒力基因的分布和耐药性调查[J].天津农学院学报,2017,24(3):55-63.Xu FX,Zhao LW,Wu YH,et al.Identification of serotype,identification of serotype,main virulence factors and investigation of drug-resistant of Vibrio parahaemolyticus strains from Tianjin strains from Tianjin[J].JTianjin Agric Coll,2017,24(3):55-63.
[13]CNAS-CL 03-A001:2018能力验证提供者认可准则在微生物领域的应用说明[Z].CNAS-CL 03-A001:2018 Guidance on the application of accreditation criteria for proficiency testing providers in the field of microbiology[Z].
[14]ISO/TS 22119 Microbiology of food and animal feeding stuffs-specific requirements and guidance for proficiency testing by interlaboratory comparison[S].
[15]CNAS-GL 002能力验证结果的统计处理和能力评价指南[S].CNAS-GL 002 Guidance on statistic treatment of proficiency testing results and performance evaluation[S].
[16]GB 4789.7-2013副溶血性弧菌检验[S].GB 4789.7-2013 Vibrio parahaemolyticus test[S].
[17]陈美玲,逄波.副溶血弧菌耐药现状及耐药机制[J].疾病监测,2018,33(5):370-375.Chen ML,Pang B.Drug resistance of Vibrio parahaemolyticus and its mechanism[J].Dis Surv,2018,33(5):370-375.
[18]强鑫华,周丽华,王萍,等.食源性腹泻副溶血弧菌分离株调查分析[J].浙江临床医学,2018,(5):971-972.Qiang XH,Zhou LH,Wang P.Investigation and analysis of food-borne diarrhea vp isolates[J].Zhejiang Clin Med J,2018,(5):971-972.
[19]杨穗珊,谢作蓉,林宝英.食品中副溶血性弧菌检测能力验证分析[J].现代食品,2018,(11):91-93.Yang SS,Xie ZR,Lin BY.Validation and analysis of the detection ability of Vibrio parahaemolyticus in food[J].Mod Food,2018,(11):91-93.
[2]Kam KM,Luey CK,Parsons MB,et al.Evaluation and validation of a PulseNet standardized pulsed-field gel electrophoresis protocol for subtyping Vibrio parahaemolyticus:An international multicenter collaborative study[J].J Clin Microbiol,2008,46(8):2766.
[3]姜大栋,李琦.一起副溶血弧菌感染食物中毒病原学检测[J].中国现代药物应用,2018,12(7):220-222.Jiang DD,Li Q.A pathogenic test of food poisoning caused by Vibrio parahaemolyticus infection[J].Chin J Mod Drug Appl,2018,12(7):220-222.
[4]施怡茹,陈洪友,卢晓芸.一起多血清型副溶血性弧菌聚集性腹泻事件的病原分析[J].检验医学与临床,2018,12(15):3662-3666.Shi YR,Chen HY,Lu XY.Pathogen analysis of a multi-serotype Vibrio parahaemolyticus aggregation diarrhea event[J].Lab Med Clin,2018,12(15):3662-3666.
[5]Croci L.Comparison of different biochemical and molecular methods for the identification of Vibrio parahaemolyticus[J].J Appl Microbiol,2010,102(1):229-237.
[6]刘秋爽.副溶血弧菌引起食物中毒调查[J].医疗装备,2018,31(1):71-72.Liu QS.Investigation of food poisoning caused by Vibrio parahaemolyticus[J].Chin J Med Dev,2018,31(1):71-72.
[7]Laskowskiarce MA,Orth K.Acanthamoeba castellanii promotes the survival of Vibrio parahaemolyticus[J].Appl Environ Microbiol,2008,74(23):7183-7188.
[8]张万标,焦辛妮,黄坚,等.2013-2015年北海市致泻性弧菌监测情况分析[J].实用预防医学,2017,24(7):865-867.Zhang WB,Jiao XN,Huang J,et al.Analysis on the monitoring of pathogenic Vibrio in beihai from 2013 to 2015[J].Pract Prev Med,2017,24(7):865-867.
[9]季霄雷,许海燕,苏婧.82株副溶血性弧菌毒力基因及耐药性分析[J].中华微生物学和免疫学杂志,2018,38(1):37-40.Ji XL,Xu HY,Su J.Analysis of virulence genes and drug resistance in 82Vibrio parahaemolyticus strains[J].Chin J Microbiol Immunol,2018,38(1):37-40.
[10]冼钰茵,余翀,阮荣勇.广州市售水产品副溶血弧菌和溶藻弧菌的耐药性评估[J].安徽农业科学,2017,45(28):74-77.Xian YY,Yu C,Ruan RY.Drug resistance of Vibrio parahemolytics and Vibrio alginolyticus in aquatic products from guangzhou retail markets[J].J Anhui Agric Sci,2017,45(28):74-77.
[11]宫春波,王朝霞,伍海燕,等.烟台海域鲜活海产品副溶血弧菌的污染概况及致病风险评价[J].青岛农业大学学报,2017,34(3):216-221.Gong CB,Wang ZX,Wu HY,et al.Pollution situation and risk assessment of Vibrio parahaemolyticus in fresh seafood in yantai sea area[J].JQingdao Agric Univ(Nat Sci),2017,34(3):216-221.
[12]徐费霞,赵良炜,吴艳辉,等.天津地区副溶血弧菌分离株血清型、部分毒力基因的分布和耐药性调查[J].天津农学院学报,2017,24(3):55-63.Xu FX,Zhao LW,Wu YH,et al.Identification of serotype,identification of serotype,main virulence factors and investigation of drug-resistant of Vibrio parahaemolyticus strains from Tianjin strains from Tianjin[J].JTianjin Agric Coll,2017,24(3):55-63.
[13]CNAS-CL 03-A001:2018能力验证提供者认可准则在微生物领域的应用说明[Z].CNAS-CL 03-A001:2018 Guidance on the application of accreditation criteria for proficiency testing providers in the field of microbiology[Z].
[14]ISO/TS 22119 Microbiology of food and animal feeding stuffs-specific requirements and guidance for proficiency testing by interlaboratory comparison[S].
[15]CNAS-GL 002能力验证结果的统计处理和能力评价指南[S].CNAS-GL 002 Guidance on statistic treatment of proficiency testing results and performance evaluation[S].
[16]GB 4789.7-2013副溶血性弧菌检验[S].GB 4789.7-2013 Vibrio parahaemolyticus test[S].
[17]陈美玲,逄波.副溶血弧菌耐药现状及耐药机制[J].疾病监测,2018,33(5):370-375.Chen ML,Pang B.Drug resistance of Vibrio parahaemolyticus and its mechanism[J].Dis Surv,2018,33(5):370-375.
[18]强鑫华,周丽华,王萍,等.食源性腹泻副溶血弧菌分离株调查分析[J].浙江临床医学,2018,(5):971-972.Qiang XH,Zhou LH,Wang P.Investigation and analysis of food-borne diarrhea vp isolates[J].Zhejiang Clin Med J,2018,(5):971-972.
[19]杨穗珊,谢作蓉,林宝英.食品中副溶血性弧菌检测能力验证分析[J].现代食品,2018,(11):91-93.Yang SS,Xie ZR,Lin BY.Validation and analysis of the detection ability of Vibrio parahaemolyticus in food[J].Mod Food,2018,(11):91-93.