克里米亞剛果出血熱病毒熒光PCR檢測(cè)試劑盒

廣州健侖生物科技有限公司

One tube multiplex for detection of Crimean Congo haemorrhagic fever virus and internal control.

單管多重檢測(cè)克里米亞剛果出血熱病毒和內(nèi)部控制。

克里米亞剛果出血熱病毒熒光PCR檢測(cè)試劑盒

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【公司名稱(chēng)】 廣州健侖生物科技有限公司
【市場(chǎng)部】    楊永漢

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7、顏色改變單位法（colour change unit,簡(jiǎn)稱(chēng)CCU）
這種方法通常用于很小，用一般的比濁法無(wú)法計(jì)數(shù)的微生物，比如支原體等，因?yàn)橹гw的液體培養(yǎng)物是*透明的，呈現(xiàn)為清亮透明紅色，因此無(wú)法用比濁法來(lái)計(jì)數(shù)，由于支原體固體培養(yǎng)很困難，用cfu法也不容易計(jì)數(shù)，因此需要用特殊的計(jì)數(shù)方法，即CCU法。它是以微生物在培養(yǎng)基中的代謝活力為指標(biāo)，來(lái)計(jì)數(shù)微生物的相對(duì)含量的，下面以解脲脲原體為例，簡(jiǎn)單介紹其操作：
（1）.取12只無(wú)菌試管，每一管裝1.8ml解脲脲原體培養(yǎng)基。
（2）.在*管加入0.2ml待測(cè)解脲脲原體菌液，充分混勻，從中吸取0.2ml加入第二管，依次類(lèi)推，10倍梯度稀釋?zhuān)恢钡絲ui末一管
（3）.于37度培養(yǎng)，以培養(yǎng)基顏色改變的zui末一管作為待測(cè)菌液的CCU，也就是支原體的zui大代謝活力，比如第六管出現(xiàn)顏色改變，他的相對(duì)濃度就是10的6次方CCU/ml.
一般來(lái)說(shuō)，比濁法和菌落計(jì)數(shù)法就可以滿足絕大多數(shù)細(xì)菌的計(jì)數(shù)，但是對(duì)支原體這樣比較特殊的微生物，用CCU法比較合適。
1.1.常量肉湯稀釋法
1.1.1.抗菌藥物貯存液制備 抗菌藥物貯存液濃度不應(yīng)低于1000μg/ml(如1280μg/ml)或10倍于zui高測(cè)定濃度。溶解度低的抗菌藥物可稍低于上述濃度。抗菌藥物直接購(gòu)自廠商或相關(guān)機(jī)構(gòu)。所需抗菌藥物溶液量或粉劑量可公式進(jìn)行計(jì)算。例如：需配制100 ml濃度為1280μg/ml的抗生素貯存液，所用抗生素為粉劑，其藥物的有效力為750μg/mg。用分析天平精確稱(chēng)取抗生素粉劑的量為182.6 mg。根據(jù)公式計(jì)算所需稀釋劑用量為：（182.6 mg×750μg/ml）/1280μg/ml=107.0ml，然后將182.6 mg抗生素粉劑溶解于107.0ml稀釋劑中。制備抗菌藥物貯存液所用的溶劑和稀釋劑見(jiàn)表5。配制好的抗菌藥物貯存液應(yīng)貯存于-60℃以下環(huán)境，保存期不超過(guò)6個(gè)月。
1.1.2.藥敏試驗(yàn)用抗菌藥物濃度范圍 根據(jù)NCCLS抗菌藥物敏感性試驗(yàn)操作標(biāo)準(zhǔn)，藥物濃度范圍應(yīng)包含耐藥、中介和敏感分界點(diǎn)值，特殊情況例外。

7, color change unit method (color change unit, referred to as CCU)
This method is generally applied to microorganisms such as mycoplasma which can not be counted by the ordinary turbidimetry because liquid cultures of mycoplasma are compley transparent and clear and bright red and thus can not be counted by turbidimetry. Because mycoplasma solid culture is very difficult to use cfu method is not easy to count, so the need for a special counting method, that CCU method. It is the microbial metabolic activity in the medium as an indicator to count the relative content of microorganisms, the following Ureaplasma urealyticum, for example, a brief description of its operation:
(1) Take 12 sterile tubes, each tube 1.8ml Ureaplasma urealyticum medium.
(2) 0.2 ml of the Ureaplasma urealyticum solution to be tested is added into the first tube, thoroughly mixed, and 0.2 ml of the Ureaplasma urealyticum is taken into the second tube, followed by a 10-fold gradient dilution until the last tube
(3) at 37 degrees Celsius, the last tube with color change medium as the test bacteria CCU, which is the maximum metabolic activity of mycoplasma, such as the sixth tube appears color change, his relative concentration is 10 6th CCU / ml.
In general, turbidimetry and colony counting method can meet the count of most bacteria, but for mycoplasma such a more specific microorganism, CCU method is more appropriate.
1.1. Constant broth dilution method
1.1.1. Antimicrobial Drug Stock Solutions Antibacterial stock solutions should be prepared at a concentration of no less than 1000 μg / ml (eg 1280 μg / ml) or 10 times the highest measured concentration. Antibacterial drugs with low solubility may be slightly lower than the above concentration. Antimicrobials are purchased directly from manufacturers or related agencies. The required amount of antibacterial solution or powder dosage can be calculated formula. For example, prepare 100 ml of antibiotic stock solution at a concentration of 1280 μg / ml. The antibiotic used is a powder and the potency of the drug is 750 μg / mg. The exact amount of antibiotic powder weighed with the analytical balance was 182.6 mg. Calculate the required amount of diluent according to the formula: (182.6 mg x 750 μg / ml) /1280μg/ml=107.0ml, then dissolve 182.6 mg of antibiotic powder in 107.0 ml of diluent. Table 5 shows the solvents and diluents used to prepare antimicrobial stock solutions. Preparation of a good antibacterial drug storage solution should be stored in the environment below -60 ℃, shelf life of not more than 6 months.
1.1.2. Range of Antimicrobials for Susceptibility Testing According to the NCCLS Antimicrobial Sensitivity Test Operating Standard, the drug concentration range should include values ??for resistance, mediation and sensitive cutoff points, with special exceptions.