培養(yǎng)基與試劑相關(guān)標(biāo)準(zhǔn)解讀
發(fā)布時間：
2022-12-24
作者：
即用型液體培養(yǎng)基公司
培養(yǎng)基與試劑相關(guān)標(biāo)準(zhǔn)解讀
微生物學(xué)檢驗(yàn)培養(yǎng)基質(zhì)量控制的意義
4789系列)都是以傳統(tǒng)微生物學(xué)培養(yǎng)為基礎(chǔ)的經(jīng)典檢驗(yàn)方法。培養(yǎng)基及相關(guān)配套試劑是微生物學(xué)常規(guī)檢驗(yàn)方法必須采用的生化試劑，培養(yǎng)基及試劑質(zhì)量會直接影響到檢驗(yàn)結(jié)果的準(zhǔn)確性，因此，必須做好培養(yǎng)基及試劑的質(zhì)量控制。涉及食品微生物學(xué)檢驗(yàn)的培養(yǎng)基種類繁多。近年來，隨著市場經(jīng)濟(jì)的發(fā)展，商品化培養(yǎng)基的應(yīng)用越來越普及?；诜N種原因，它們的配方、用途、使用方法、保存方式各不相同，為實(shí)驗(yàn)室質(zhì)量控制帶來新問題?！ ?/span>
目前，作為我國現(xiàn)行食品安全標(biāo)準(zhǔn)體系配套的強(qiáng)制性標(biāo)準(zhǔn)《食品安全國家標(biāo)準(zhǔn)食品微生物學(xué)檢驗(yàn)》(GB1538.2—2016《培養(yǎng)基制備指南第2部分：培養(yǎng)基性能測試實(shí)用指南》進(jìn)行[2，3]，但是，這些相關(guān)標(biāo)準(zhǔn)或規(guī)定受限于應(yīng)用領(lǐng)域，作為微生物學(xué)檢驗(yàn)培養(yǎng)基和試劑的質(zhì)量控制依據(jù)也不完備，多數(shù)商品化培養(yǎng)基生產(chǎn)企業(yè)也未據(jù)此作為質(zhì)量控制標(biāo)準(zhǔn)，而檢驗(yàn)機(jī)構(gòu)對自行配制的培養(yǎng)基進(jìn)行質(zhì)量控制的情況更少，導(dǎo)致培養(yǎng)基和試劑質(zhì)量參差不齊。據(jù)以往相關(guān)調(diào)查，國產(chǎn)培養(yǎng)基的質(zhì)量均有嚴(yán)重問題[4]。中國國家認(rèn)證認(rèn)可監(jiān)督管理委員會網(wǎng)站通報(bào)的CNAS實(shí)驗(yàn)室現(xiàn)場評審在《服務(wù)和供應(yīng)品的采購》這個環(huán)節(jié)的常見不符合項(xiàng)最多，主要是：未建立培養(yǎng)基和試劑質(zhì)量控制程序、不能提供關(guān)鍵培養(yǎng)基技術(shù)性驗(yàn)收評估記錄和不能提供實(shí)驗(yàn)室用水符合性評價記錄，說明多數(shù)檢驗(yàn)機(jī)構(gòu)對于培養(yǎng)基和試劑的相關(guān)標(biāo)準(zhǔn)和規(guī)定的理解、執(zhí)行都存在偏差。第1部分：實(shí)驗(yàn)室培養(yǎng)基制備質(zhì)量保證通則》和SN/T1538.1—2016《培養(yǎng)基制備指南1133制定的SN/T11133—2014《食品、動物飼料和水的微生物學(xué)—營養(yǎng)培養(yǎng)基的制備、生產(chǎn)、存儲和性能測試》或修改采用ISO1 現(xiàn)行的相關(guān)微生物學(xué)檢驗(yàn)用培養(yǎng)基質(zhì)量控制的標(biāo)準(zhǔn)或規(guī)定，除了2013版標(biāo)準(zhǔn)之外，CNAS—CL09：2013《檢測和校準(zhǔn)實(shí)驗(yàn)室能力認(rèn)可準(zhǔn)則在微生物檢測領(lǐng)域的應(yīng)用說明》規(guī)定實(shí)驗(yàn)室對于培養(yǎng)基和試劑的驗(yàn)收可參考ISO。
培養(yǎng)基和試劑質(zhì)量控制的要求。
在中國國家認(rèn)證認(rèn)可監(jiān)督管理委員會發(fā)布的《檢驗(yàn)檢測機(jī)構(gòu)資質(zhì)認(rèn)定評審準(zhǔn)則》4.5.6條款中，規(guī)定“檢驗(yàn)檢測機(jī)構(gòu)應(yīng)建立和保持選擇和購買對檢驗(yàn)檢測質(zhì)量有影響的服務(wù)和供應(yīng)品的程序。明確服務(wù)、供應(yīng)品、試劑、消耗材料的購買、驗(yàn)收、存儲的要求”，就是要求“在中華人民共和國境內(nèi)，向社會出具具有證明作用的數(shù)據(jù)、結(jié)果的檢驗(yàn)檢測機(jī)構(gòu)”為保證采購物品和相關(guān)服務(wù)的質(zhì)量，應(yīng)當(dāng)按制定的程序?qū)υ噭┻M(jìn)行有效的控制和管理，以保證檢驗(yàn)檢測結(jié)果的質(zhì)量[5]。雖然文件有規(guī)定，但是，具體如何執(zhí)行，需要檢驗(yàn)機(jī)構(gòu)結(jié)合自身實(shí)際情況制定程序性文件。2013版標(biāo)準(zhǔn)只是食品安全國家標(biāo)準(zhǔn)，適用范圍僅限于食品微生物學(xué)檢驗(yàn)領(lǐng)域，而目前其他微生物學(xué)檢驗(yàn)領(lǐng)域并沒有適用的培養(yǎng)基和試劑質(zhì)量控制標(biāo)準(zhǔn)，所以，建議所有涉及微生物學(xué)檢驗(yàn)領(lǐng)域的檢驗(yàn)機(jī)構(gòu)都執(zhí)行或參照執(zhí)行2013版標(biāo)準(zhǔn)，制定適合本實(shí)驗(yàn)室的培養(yǎng)基和試劑質(zhì)量控制程序性文件。
2013版標(biāo)準(zhǔn)的具體要求和存在問題
術(shù)語和定義
11133分為6類有所不同，例如，將選擇性計(jì)數(shù)培養(yǎng)基分為選擇性固體計(jì)數(shù)培養(yǎng)基和選擇性液體計(jì)數(shù)培養(yǎng)基，明顯就是畫蛇添足?！ ”菊鹿?jié)主要缺陷是定義描述不到位和前后術(shù)語不一致。同時，遺漏了診斷血清、測試紙片、生化鑒定試劑盒等常用微生物檢驗(yàn)試劑。另外，標(biāo)準(zhǔn)將培養(yǎng)基和試劑分為11類，分類方法與ISO
缺少“設(shè)備和儀器”
作為一個既有檢驗(yàn)方法又有判定依據(jù)的標(biāo)準(zhǔn)，缺少對于設(shè)備和儀器的規(guī)定，是不可想象的。
培養(yǎng)基及試劑質(zhì)量保證
這個章節(jié)分為證明文件、貯存、培養(yǎng)基的實(shí)驗(yàn)室制備、培養(yǎng)基的使用、培養(yǎng)基的棄置五個部分。建議檢驗(yàn)機(jī)構(gòu)按照這個框架規(guī)定培養(yǎng)基和試劑的質(zhì)量控制程序。這個章節(jié)存在的問題，主要是對培養(yǎng)基保質(zhì)期規(guī)定太松、配制用水電阻率錯誤、pH測定溫度不適用、培養(yǎng)基的棄置可操作性不強(qiáng)，而目前現(xiàn)場評審時，實(shí)驗(yàn)室被核查到的常見錯誤，除了培養(yǎng)基配制原始記錄信息不全這個通病外，幾乎所有實(shí)驗(yàn)室都沒有做到“傾注瓊脂量達(dá)到3mm以上厚度”，屬于未認(rèn)真學(xué)習(xí)標(biāo)準(zhǔn)規(guī)定。
質(zhì)控菌株的保藏及使用
19489—2008等其他實(shí)驗(yàn)室質(zhì)量控制管理規(guī)定要求不一致[2，6，7]，容易理解錯誤?？倓t》、GB食品微生物學(xué)檢驗(yàn)4789.1—2016《食品安全國家標(biāo)準(zhǔn) 這個章節(jié)，除了使用術(shù)語與第二章規(guī)定不一致外，最重要的問題是描述和規(guī)定不夠詳盡，也未考慮生物安全相關(guān)標(biāo)準(zhǔn)要求，因此，與CNAS—CL09：2013、GB
培養(yǎng)基和試劑的質(zhì)量要求
1.基本要求
本條規(guī)定供應(yīng)商和制備者應(yīng)進(jìn)行培養(yǎng)基和試劑的20℃～25℃的pH和緩沖能力特性評估，前者與標(biāo)準(zhǔn)全文都在25℃測定pH的規(guī)定不符，后者則是全文都沒有出現(xiàn)如何檢驗(yàn)、如何判定的描述。
2.微生物學(xué)要求
概論規(guī)定實(shí)驗(yàn)室應(yīng)“制定驗(yàn)收程序，如需進(jìn)行驗(yàn)證，可按6.2執(zhí)行”。對此條款，目前，多數(shù)檢驗(yàn)機(jī)構(gòu)并未理解，除了沒有制定明確的培養(yǎng)基和試劑驗(yàn)收程序外，也不知道哪些培養(yǎng)基和試劑需要進(jìn)行驗(yàn)證。其實(shí)，就是有第三方檢驗(yàn)機(jī)構(gòu)檢驗(yàn)報(bào)告判定“達(dá)到附錄D質(zhì)量控制標(biāo)準(zhǔn)的要求”的，以及對檢驗(yàn)結(jié)果沒有直接影響的培養(yǎng)基和試劑，不需要驗(yàn)證。在“5.2.3.2測試菌株”條，使用了與第四章“質(zhì)控菌株”不一樣的術(shù)語。這個條款中，規(guī)定了根據(jù)培養(yǎng)基和試劑類型的不同，采用的測試菌株數(shù)量。在現(xiàn)場評審時，常見評審員要求實(shí)驗(yàn)室必須使用附錄E里面的全部菌株進(jìn)行測試或?qū)嶒?yàn)室僅用一株菌株進(jìn)行選擇性培養(yǎng)基測試等情況，都屬于對標(biāo)準(zhǔn)理解錯誤。
培養(yǎng)基和試劑性能測試方法
總體來說，2016版標(biāo)準(zhǔn)考慮到我國中小型實(shí)驗(yàn)室進(jìn)行全部培養(yǎng)基和試劑驗(yàn)證的工作量，并未一味照搬國際標(biāo)準(zhǔn)的要求，人性化規(guī)定了實(shí)驗(yàn)室可以使用半定量和定性測試方法驗(yàn)證培養(yǎng)基和試劑，有利于實(shí)驗(yàn)室執(zhí)行并開展。不足之處是，本章節(jié)文字、語法及描述性錯誤太多，影響了標(biāo)準(zhǔn)的實(shí)施效果。
測試結(jié)果的記錄
7.1條要求“培養(yǎng)基制造商或供應(yīng)商應(yīng)按客戶的要求提供培養(yǎng)基常規(guī)信息和相關(guān)測試菌株生長特性信息。”與3.1.1條“生產(chǎn)企業(yè)提供的文件”屬于重復(fù)規(guī)定。
7.2條要求“按照質(zhì)量體系的要求，對所有培養(yǎng)基性能測試的數(shù)據(jù)歸檔并在有效期內(nèi)進(jìn)行適當(dāng)?shù)谋４妗?rdquo;實(shí)際上，任何質(zhì)量控制體系都沒有規(guī)定必須對所有培養(yǎng)基進(jìn)行性能測試，用于輔助檢驗(yàn)用的培養(yǎng)基，如，菌種保存、實(shí)驗(yàn)室內(nèi)部環(huán)境監(jiān)控的那些培養(yǎng)基并不強(qiáng)制進(jìn)行性能測試，只要對關(guān)鍵性培養(yǎng)基和試劑進(jìn)行驗(yàn)證即可。
Standard interpretation of medium and reagent
Significance of microbiological examination of medium quality control
Series 4789) are all classical test methods based on traditional microbial culture. Medium and related auxiliary reagent is routine microbiological testing method must be used biochemical reagents, culture medium and reagent quality will directly affect the accuracy of test results, therefore, culture medium and the quality control of the reagents must be done. There are many kinds of media involved in food microbiological testing. In recent years, with the development of market economy, the application of commercial media is more and more popular. For various reasons, their formulations, USES, use methods and preservation methods are different, which bring new problems to laboratory quality control.
Currently, as a compulsory standard in China's current food safety standard system, food safety national standard food microbiology test (gb1538.2-2016) Medium performance test and practical guide to [2, 3], however, the relevant standards or provisions are limited by application field, as the quality control of microbiology test reagent and medium basis is not complete, most of the commercialization of media production enterprises is not as a quality control standard, accordingly the inspection institution to preparation of culture medium for quality control of less, leading to medium and reagents are of variable quality. According to previous relevant surveys, the quality of domestic media has serious problems [4]. The supervision and administration of the China national accreditation committee website report CNAS lab site assessment in the procurement of services and supplies at this stage the most common nonconformities, mainly: medium and reagent quality control program has not been established, is unable to provide the key medium technical acceptance evaluation records and cannot provide laboratory water conformity assessment records, shows that most of the inspection institution for the understanding of culture medium and the related standards and regulations of the reagents, execution, there is a deviation. Part 1: general principles for quality assurance laboratory culture medium preparation and SN/T1538.1-2016 "medium preparation guidelines set by the 1133 SN/T11133-2014" food, animal feed and water microbiology, nutrient medium preparation, production, storage, and the performance test, or modify the ISO1 current microbiology related media quality control standards or regulations, in addition to the 2013 version of the standard, CNAS - CL09: In 2013, the application description of the laboratory accreditation criteria for testing and calibration in the field of microbiological detection stipulated that the laboratory's acceptance of media and reagents could refer to ISO.
Quality control requirements for media and reagents.
Released in China national accreditation supervision and administration commission of the assessment criteria testing institutions aptitude 4.5.6 terms, provisions of "the examination institution shall establish and maintain affects the examination quality to select and purchase services and supplies. Specific services, supplies, reagents, consumable material purchase, acceptance, storage requirements ", is required to the society "within the territory of the People's Republic of China, has the effective evidence issued by the data, the results of the examination institutions" to ensure the quality of procurement of goods and related services, shall be formulated according to the procedure of reagents for effective control and management, to ensure the quality of the examination results [5]. Although there are provisions in the documents, it is necessary for the inspection authorities to formulate procedural documents according to their actual conditions. 2013 version of the standard is the national food safety standards, the applicable scope is limited to food microbiology test field, and the other field of microbiology test was not applicable medium and reagent quality control standards, therefore, recommended that all involved in the field of microbiology test inspection institutions are executed at or around the 2013 version of the standard, suitable for the laboratory reagent and medium quality control programmatic document.
Specific requirements and existing problems of the 2013 edition standard
Terms and definitions
For example, the selective count medium was divided into the selective solid count medium and the selective liquid count medium. The main drawback of this chapter is that the definition description is not in place and the terms are inconsistent. At the same time, the diagnostic serum, test paper, biochemical identification kit and other commonly used microbial test reagents were omitted. In addition, the standard divides media and reagents into 11 categories, classification methods and ISO
Lack of "equipment and equipment"
As a standard that has both inspection method and judgment basis, it is impossible to imagine the lack of provisions on equipment and instruments.
Medium and reagent quality assurance
This chapter is divided into five parts: proof document, storage, laboratory preparation of medium, use of medium and discard of medium. It is recommended that inspection bodies establish quality control procedures for media and reagents in accordance with this framework. Problems existing in this chapter, mainly for medium shelf life is too loose, mixed water resistivity error, shall not apply to the pH measurement temperature and medium disposed operability is not strong, and the site assessment, laboratory verification to common errors, in addition to the culture medium preparation records the information is not complete and the common fault, almost all the laboratory do not "pour into AGAR volume reached more than 3 mm thickness, is not seriously learning standards.
Storage and use of quality control strains
Other laboratory quality control regulations, such as 19489-2008, have inconsistent requirements [2,6,7] and are easy to understand and make mistakes. General rules ", GB food microbiology test 4789.1-2016 "the national food safety standards of this section, in addition to use the term inconsistent with the provisions of the second chapter, the most important problem is not enough detailed descriptions and regulation, also did not consider biological safety standard requirements, therefore, with CNAS - CL09:2013, GB
Quality requirements for medium and reagent
1. Basic requirements
Should be carried out the provisions of the supplier and the preparation of culture medium and reagents of 20 ℃ to 25 ℃ and buffering capacity of the pH value of the property evaluation, the former with the standard does not conform to that stipulated in full all at 25 ℃ was developed for the determination of pH, the latter is the full text is not how to test, how to determine the description.
2. Microbiological requirements
The introduction stipulates that the laboratory should "develop acceptance procedures, and if verification is required, it can be carried out in accordance with 6.2". At present, most inspection institutions do not understand this provision, and they do not know which media and reagents need to be verified except that there is no clear culture medium and reagent acceptance procedure. Actually, is the third party inspection agency inspection report to judge "to achieve the requirement of the appendix D quality control standard", and not directly influence the result of the test medium and reagents, don't need to test and verify. In the section "5.2.3.2 test strains", a different term is used than that used in chapter 4, "quality control strains". This provision specifies the number of test strains to be used depending on the type of medium and reagent. On site review, common judges require laboratory must use the inside of the appendix E all the strains tested or lab in only one strains tested selective medium, etc., all belong to understanding standard error.
Test method for performance of culture medium and reagent
2016 version of the standard, in general, considering our country small and medium-sized lab for all media and reagents to verify the amount of work, not blindly copy the requirements of international standard, humanized rules the lab can use half quantitative and qualitative test method validation of reagent and medium, is advantageous to the laboratory to execute and carry out. The disadvantage is that there are too many text, grammar and descriptive errors in this chapter, which affect the implementation effect of the standard.
Records of test results
7.1 requirements: "the medium manufacturer or supplier shall provide the conventional information of the medium and the relevant information on the growth characteristics of the tested strains as required by the customer." And 3.1.1 "documents provided by production enterprises" are duplicated.
7.2 "according to the requirements of the quality system, the data of all performance tests of the medium shall be filed and properly kept within the validity period." In fact, did not set any quality control system must be conducted for all media performance test, used for auxiliary test in culture medium, such as, species conservation, internal environmental monitoring laboratory performance testing of the medium is not mandatory, as long as the key media and reagents for validation.