犬弓形蟲定量檢測卡TOXO

廣州健侖生物科技有限公司

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【犬弓形蟲定量檢測卡TOXO】

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【公司名稱】 廣州健侖生物科技有限公司

【市 場 部】    楊永漢

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【騰訊Q Q】 2042552662

【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號二期2幢101-103室

1.1.5.稀釋抗菌藥物的制備及菌液接種 取無菌試管（13×100mm）13支，排成一排，除第1管加入1.6mlMH肉湯外，其余每管加入MH肉湯1ml，在第1管加入抗菌藥物原液（如1280μg/ml） 0.4ml混勻，然后吸取1ml至第2管，混勻后再吸取1ml至第3管，如此連續(xù)倍比稀釋至第11管，并從第11管中吸取1ml棄去，第12管為不含藥物的生長對照。此時各管藥物濃度依次為256、128、64、32、16、8、4、2、1、0.5、0.25μg/ml。然后在每管內(nèi)加入上述制備好的接種物各1ml，使每管zui終菌液濃度約為5×105CFU/ml。第1管至第11管藥物濃度分別為128、64、32、16、8、4、2、1、05、0.25、0.125μg/ml。
1.1.6.孵育 將接種好的稀釋管塞好塞子，置35℃普通空氣孵箱中孵育16~20h；嗜血桿菌和鏈球菌在普通空氣孵箱中孵育20~24h；對可能的耐甲氧西林葡萄球菌和耐萬古霉素腸球菌應(yīng)持續(xù)孵育滿24h。
1.1.7.結(jié)果判斷與解釋 在讀取和報告所測試菌株的MIC前，應(yīng)檢查生長對照管的細菌生長情況是否良好，同時還應(yīng)檢查接種物的傳代培養(yǎng)情況以確定其是否污染，質(zhì)控菌株的MIC值是否處于質(zhì)控范圍。以肉眼觀察，藥物zui低濃度管無細菌生長者，即為受試菌的MIC。甲氧芐胺嘧啶或磺胺藥物的肉湯稀釋法終點判斷，與陽性生長對照管比較抑制80%細菌生長管藥物濃度為受試菌MIC。
根據(jù)NCCLS推薦的分界點值標(biāo)準(zhǔn)，判斷耐藥（resistant, R）、敏感(susceptible, S)或中介（intermediate, I）。S表示被測菌株所引起的感染可以用該抗菌藥物的常用劑量治療有效，禁忌癥除外。R指該菌不能被抗菌藥物的常用劑量在組織液內(nèi)或血液中所達到的濃度所抑制，或?qū)儆诰哂刑囟退帣C理（如β-內(nèi)酰胺酶），所以臨床治療效果不佳。I是指MIC接近藥物的血液或組織液濃度，療效低于敏感菌。還表示被測菌株可以通過提高劑量（如β-內(nèi)酰胺類藥物）被抑制，或在藥物生理性濃集的部位（如尿液）被抑制。另外，中介還作為“緩沖域”，以防止由微小的技術(shù)因素失控，所導(dǎo)致較大的錯誤解釋。
1.2.微量肉湯稀釋法 
1.2.1.抗菌藥物和培養(yǎng)基制備 同常量肉湯稀釋法。
1.2.2.MIC板制備 無菌操作，將倍比稀釋后不同濃度的抗菌藥物溶液分別加到滅菌的96孔聚苯乙烯板中，第1至第11孔加藥液，每孔10μl，第12孔不加藥作為生長對照，冰凍干燥后密封，-20℃以下保存?zhèn)溆谩?/p>

1.1.5. Preparation of diluted antibacterial drugs and bacterial inoculum take sterile tube (13 × 100mm) 13, arranged in a row, in addition to the first tube to join 1.6mlM broth, the rest of each tube was added MH broth 1ml, In the first tube by adding antibacterial stock solution (such as 1280μg / ml) 0.4ml mix, then draw 1ml to the second tube, mix and then draw 1ml to the third tube, so continuous serial dilution to 11 tubes, and from Pipette 1ml discard the first 11, the first 12 tubes for the drug-free growth control. At this point the drug concentration in each tube were 256,128,64,32,16,8,4,2,1,0.5,0.25μg / ml. Then add 1ml each of the prepared inoculum in each tube so that the final bacterial concentration of each tube is about 5 × 10 5 CFU / ml. The first to the eleventh tube drug concentrations were 128,64,32,16,8,4,2,1,05,0.25,0.125μg / ml.
1.1.6. Incubation Inoculation of a good dilution pipe plug plug, set 35 ℃ normal air incubator incubated 16 ~ 20h; Haemophilus and Streptococcus incubated in an ordinary air incubator 20 ~ 24h; the possible resistance to a Staphylococcus aureus and vancomycin-resistant enterococci should be incubated for 24 hours.
1.1.7. Judgment and interpretation of results Before reading and reporting the MIC of the tested strains, the growth of the bacteria in the growth control should be checked for good bacterial growth, and the inoculum culture should be checked for contamination, control The MIC value of the strain is in the control range. Observed with the naked eye, the minimum concentration of drug-free bacterial growth, that is, the MIC of the test bacteria. Trimethoprim or sulfa drug broth dilution method to determine the endpoint, compared with the positive growth control tube 80% inhibition of bacterial growth tube drug concentration MIC for the test bacteria.
According to the NCCLS recommended cut-off value criteria, it is judged resistant, susceptible, intermediate or I. S indicates that the infection caused by the tested strain can be treated with the usual dose of the antimicrobial agent except contraindications. R means that the bacteria can not be the usual dose of antimicrobial agents in the tissue fluid or blood concentrations reached by inhibition, or belong to a specific resistance mechanism (such as β-lactamase), so the clinical curative effect is not good. I refers to the MIC close to the drug blood or tissue fluid concentration, less effective than sensitive bacteria. It also indicates that the tested strain can be inhibited by increasing doses (such as beta-lactam) or at sites where the drug is physiologically concentrated (such as urine). In addition, the intermediary also acts as a "buffer zone" to prevent large technical errors from being overseen by minor technical considerations.
1.2. Micro broth dilution method
1.2.1. Antibacterial drugs and medium preparation with the same constant broth dilution method.
1.2.2.MIC plate preparation of aseptic operation, the dilution ratio of different concentrations of antibacterial drug solution were added to sterilized 96-well polystyrene plates, the first 11 holes plus liquid, each hole 10μl, The first 12 wells without drug as a growth control, freeze-dried and sealed, -20 ℃ the following reserve.