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一、大腸桿菌大腸菌群微生物檢測板產品介紹
1、原理及適用范圍
大(da)(da)(da)(da)腸(chang)(chang)(chang)桿(gan)(gan)菌(jun)(jun)(jun)(jun)(Escherichia coli)和(he)(he)(he)大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun)(Coliform)都是存(cun)在于(yu)溫血(xue)動物腸(chang)(chang)(chang)道(dao)和(he)(he)(he)糞(fen)便(bian)的(de)細菌(jun)(jun)(jun)(jun),是條件性(xing)致病菌(jun)(jun)(jun)(jun),主(zhu)要(yao)造(zao)成(cheng)水(shui)體污(wu)染,也可(ke)能引起食(shi)物中毒的(de)發生。大(da)(da)(da)(da)腸(chang)(chang)(chang)桿(gan)(gan)菌(jun)(jun)(jun)(jun)大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun)檢(jian)(jian)測(ce)板是一(yi)種預先(xian)制備好的(de)一(yi)次性(xing)培(pei)養基產(chan)品(pin),可(ke)以同時檢(jian)(jian)測(ce)大(da)(da)(da)(da)腸(chang)(chang)(chang)桿(gan)(gan)菌(jun)(jun)(jun)(jun)和(he)(he)(he)大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun),含(han)有(you)大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)選擇性(xing)培(pei)養基、冷水(shui)可(ke)溶性(xing)的(de)吸(xi)水(shui)凝膠和(he)(he)(he)半乳糖(tang)苷酶(GAL)和(he)(he)(he)葡萄糖(tang)醛酸酶(GUD)指示劑(ji)。大(da)(da)(da)(da)腸(chang)(chang)(chang)桿(gan)(gan)菌(jun)(jun)(jun)(jun)顯藍(lan)色(se),其他大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun)顯紅色(se),藍(lan)點加上紅點為總的(de)大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun)數(shu)。本產(chan)品(pin)適(shi)合(he)于(yu)食(shi)品(pin)及原料(liao)中大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun)的(de)計(ji)(ji)(ji)數(shu),如消(xiao)毒牛乳、冷飲和(he)(he)(he)調味品(pin)等樣品(pin)的(de)檢(jian)(jian)測(ce),也可(ke)用于(yu)表面的(de)衛生檢(jian)(jian)測(ce)。 執行(xing)標(biao)準:中華(hua)人民(min)共(gong)(gong)和(he)(he)(he)國國家(jia)標(biao)準《食(shi)品(pin)安全國家(jia)標(biao)準 食(shi)品(pin)微(wei)(wei)生物檢(jian)(jian)驗 大(da)(da)(da)(da)腸(chang)(chang)(chang)埃希氏菌(jun)(jun)(jun)(jun)計(ji)(ji)(ji)數(shu)》(GB 4789.38);中華(hua)人民(min)共(gong)(gong)和(he)(he)(he)國出入境檢(jian)(jian)驗檢(jian)(jian)疫行(xing)業(ye)標(biao)準《食(shi)品(pin)中大(da)(da)(da)(da)腸(chang)(chang)(chang)菌(jun)(jun)(jun)(jun)群(qun)和(he)(he)(he)大(da)(da)(da)(da)腸(chang)(chang)(chang)桿(gan)(gan)菌(jun)(jun)(jun)(jun)快速(su)計(ji)(ji)(ji)數(shu)法——PetrifilmTM 檢(jian)(jian)測(ce)板法》(SN/T 1896)。
2、大腸桿菌大腸菌群微生物檢測板操作方法
2.1 樣品處理:取(qu)樣品 25 mL(g)放入含有225 mL滅菌磷酸緩沖液(ye)(或生理鹽水(shui))的(de)(de)取(qu)樣罐(guan)或均質袋內,制成1:10的(de)(de)樣品勻液(ye),根據(ju)樣品污染程度及檢驗需(xu)要,可(ke)進一步制成10倍(bei)遞增的(de)(de)樣品稀釋液(ye)。
2.2 接種:一般(ban)食品(pin)選2~3個(ge)稀釋(shi)度進(jin)(jin)行檢(jian)測(ce)(ce),含(han)菌(jun)量少(shao)的液(ye)(ye)體樣(yang)品(pin)(如飲用純水(shui)和礦(kuang)泉水(shui)等)可直(zhi)接吸(xi)(xi)(xi)取原液(ye)(ye)進(jin)(jin)行檢(jian)測(ce)(ce)。揭開大腸菌(jun)群檢(jian)測(ce)(ce)板(ban)(ban)上(shang)(shang)層(ceng)膜(mo),用無菌(jun)吸(xi)(xi)(xi)管吸(xi)(xi)(xi)取1 mL樣(yang)品(pin)勻(yun)液(ye)(ye)滴(di)加到檢(jian)測(ce)(ce)板(ban)(ban)內,迅(xun)速貼(tie)好(hao)上(shang)(shang)層(ceng)膜(mo)并水(shui)平晃動檢(jian)測(ce)(ce)板(ban)(ban),讓樣(yang)品(pin)勻(yun)液(ye)(ye)均(jun)勻(yun)吸(xi)(xi)(xi)附在濾紙上(shang)(shang),靜置(zhi)10 s左右,待(dai)樣(yang)品(pin)勻(yun)液(ye)(ye)被吸(xi)(xi)(xi)附在濾紙上(shang)(shang)。每個(ge)稀釋(shi)度接種兩片(pian)。做好(hao)空白陰性對照。
2.3 培(pei)養:將檢測(ce)板疊在一(yi)起,倒置放(fang)于培(pei)養箱內。36℃±1℃,培(pei)養15~24h。
3、結果判讀
培養后(hou)大腸(chang)(chang)桿(gan)菌(jun)顯藍(lan)色,其他大腸(chang)(chang)菌(jun)群(qun)顯紅(hong)色,藍(lan)點(dian)加(jia)上紅(hong)點(dian)為總(zong)的大腸(chang)(chang)菌(jun)群(qun)數。選(xuan)擇有(you)(you)陽性(xing)菌(jun)落(luo)的較(jiao)高稀(xi)釋度的檢測板進行計數,然后(hou)乘以稀(xi)釋倍數即(ji)為樣品中(zhong)每毫升(sheng)(克)含有(you)(you)大腸(chang)(chang)菌(jun)群(qun)數。
4、附加說明
4.1 我國的(de)食品衛(wei)生標(biao)準大(da)多都以大(da)腸菌群(Coliform)作為檢測指標(biao),而歐盟等許多國家(jia)則以大(da)腸桿菌(E.coli)作為檢測對象(xiang),因(yin)此(ci),本產品對于出口(kou)食品和航(hang)空配餐等企業具有更廣泛的(de)應用前景。
4.2 樣品勻(yun)液的pH值應在6.5~7.5之間(jian),需要時分別(bie)用1mol/L NaOH或(huo)者1mol/L HCl調節。
4.3 大(da)腸桿(gan)菌(jun)(jun)大(da)腸菌(jun)(jun)群檢測板對純菌(jun)(jun)的檢測靈敏(min)度可達(da)100 cfu/mL。
4.4 一般食(shi)品對于大腸菌(jun)群的要求都比較嚴格,如果(guo)檢測板上(shang)有成(cheng)片的紅點(dian),或者中央沒(mei)有紅點(dian),但邊緣有很多紅點(dian),定是已經嚴重(zhong)超標,建議按(an)“多不可(ke)計”來報告。
4.5注意使用過的檢測板上帶有活菌(jun),需(xu)及時按照(zhao)生物(wu)安全廢(fei)棄物(wu)處理(li)原(yuan)則進行處理(li)。