This document specifies a horizontal method for the detection of enteropathogenic Vibrio<\/i> spp., which causes human illness in or via the intestinal tract. The species detectable by the methods specified include Vibrio parahaemolyticus, Vibrio cholerae<\/i> and Vibrio vulnificus<\/i>.<\/p>\n
It is applicable to the following:<\/p>\n
products intended for human consumption and the feeding of animals;<\/p>\n<\/li>\n
environmental samples in the area of food production and food handling.<\/p>\n<\/li>\n<\/ul>\n
This method may not be appropriate in every detail for certain products (see Introduction).<\/p>\n<\/div>\n
The World Health Organization (WHO) has identified that V. parahaemolyticus, V. cholerae<\/i> and V. vulnificus<\/i> are the major food-borne Vibrio<\/i> spp. However, the method in this document can also be appropriate for the identification of other Vibrio<\/i> spp. causing illness in humans. [1]<\/sup> <\/span><\/p>\n<\/div>\n Microbiology of the food chain. Horizontal method for the determination of Vibrio spp. – Detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus<\/b><\/p>\nPDF Catalog<\/h4>\n
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\n PDF Pages<\/th>\n PDF Title<\/th>\n<\/tr>\n \n 2<\/td>\n National foreword <\/td>\n<\/tr>\n \n 4<\/td>\n European foreword
Endorsement notice <\/td>\n<\/tr>\n\n 7<\/td>\n Foreword <\/td>\n<\/tr>\n \n 8<\/td>\n Introduction <\/td>\n<\/tr>\n \n 9<\/td>\n 1 Scope
2 Normative references <\/td>\n<\/tr>\n\n 10<\/td>\n 3 Terms and definitions
4 Principle
4.1 General
4.2 Primary enrichment in a liquid selective medium <\/td>\n<\/tr>\n\n 11<\/td>\n 4.3 Secondary enrichment in a liquid selective medium
4.4 Isolation and identification
4.5 Confirmation
5 Culture media and reagents <\/td>\n<\/tr>\n\n 12<\/td>\n 5.1 Enrichment medium: alkaline saline peptone water (ASPW)
5.2 Solid selective isolation media
5.2.1 First medium: thiosulphate, citrate, bile and sucrose agar medium (TCBS)
5.2.2 Second medium
5.3 Saline nutrient agar (SNA)
5.4 Reagent for detection of oxidase
5.5 Biochemical tests
5.5.1 L-lysine decarboxylase saline medium (LDC)
5.5.2 Arginine dihydrolase saline medium (ADH) <\/td>\n<\/tr>\n\n 13<\/td>\n 5.5.3 Reagent for detection of \u03b2-galactosidase
5.5.4 Saline medium for detection of indole
5.5.5 Saline peptone waters
5.5.6 Sodium chloride solution
5.6 PCR
5.6.1 Tris acetate EDTA buffer (TAE) (or a buffer allowing similar performance for the purpose)
5.6.2 Mastermix
5.6.3 Primers and probes
5.6.4 Positive control material <\/td>\n<\/tr>\n\n 14<\/td>\n 5.6.5 Negative extraction control
6 Equipment and consumables
7 Sampling
8 Preparation of the test sample <\/td>\n<\/tr>\n\n 15<\/td>\n 9 Procedure (See Figure\u00a0A.1)
9.1 Test portion and initial suspension
9.2 Primary selective enrichment
9.3 Secondary selective enrichment <\/td>\n<\/tr>\n\n 16<\/td>\n 9.4 Isolation and identification
9.5 Confirmation
9.5.1 General <\/td>\n<\/tr>\n\n 17<\/td>\n 9.5.2 Selection of colonies for confirmation and preparation of pure cultures
9.5.3 Tests for presumptive identification <\/td>\n<\/tr>\n\n 18<\/td>\n 9.5.4 Biochemical confirmation <\/td>\n<\/tr>\n \n 20<\/td>\n 9.5.5 PCR confirmation
9.5.6 DNA extraction
9.5.7 Conventional PCR <\/td>\n<\/tr>\n\n 21<\/td>\n 9.5.8 Real-time PCR
10 Expression of results
11 Performance characteristics of the method
11.1 Interlaboratory study <\/td>\n<\/tr>\n\n 22<\/td>\n 11.2 Sensitivity
11.3 Specificity
11.4 LOD50
12 Test report <\/td>\n<\/tr>\n\n 23<\/td>\n Annex\u00a0A (normative) Diagram of procedure <\/td>\n<\/tr>\n \n 25<\/td>\n Annex\u00a0B (normative) Composition and preparation of the culture media and reagents <\/td>\n<\/tr>\n \n 31<\/td>\n Annex\u00a0C (informative) Conventional PCR for the detection of Vibrio parahaemolyticus, thermostable direct haemolysin (tdh) and thermostable direct related haemolysin (trh) genes, Vibrio cholerae and Vibrio vulnificus <\/td>\n<\/tr>\n \n 35<\/td>\n Annex\u00a0D (informative) Real-time PCR for the detection of Vibrio parahaemolyticus, thermostable direct haemolysin gene (tdh) and Vibrio vulnificus <\/td>\n<\/tr>\n \n 37<\/td>\n Annex\u00a0E (informative) Results of an interlaboratory study <\/td>\n<\/tr>\n \n 40<\/td>\n Bibliography <\/td>\n<\/tr>\n<\/table>\n","protected":false},"excerpt":{"rendered":" \n\n
\n Published By<\/td>\n Publication Date<\/td>\n Number of Pages<\/td>\n<\/tr>\n \n BSI<\/b><\/a><\/td>\n 2017<\/td>\n 42<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n","protected":false},"featured_media":341519,"template":"","meta":{"rank_math_lock_modified_date":false,"ep_exclude_from_search":false},"product_cat":[124,2641],"product_tag":[],"class_list":{"0":"post-341512","1":"product","2":"type-product","3":"status-publish","4":"has-post-thumbnail","6":"product_cat-07-100-30","7":"product_cat-bsi","9":"first","10":"instock","11":"sold-individually","12":"shipping-taxable","13":"purchasable","14":"product-type-simple"},"_links":{"self":[{"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/product\/341512","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/product"}],"about":[{"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/types\/product"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/media\/341519"}],"wp:attachment":[{"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/media?parent=341512"}],"wp:term":[{"taxonomy":"product_cat","embeddable":true,"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/product_cat?post=341512"},{"taxonomy":"product_tag","embeddable":true,"href":"https:\/\/pdfstandards.shop\/wp-json\/wp\/v2\/product_tag?post=341512"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}