病毒性脑膜炎5联荧光PCR检测试剂盒

病毒性脑膜炎5联荧光PCR检测试剂盒

广州健仑生物科技有限公司

准备使用lyo master混合物（每个8孔条）检测单纯疱疹病毒1型，单纯疱疹病毒2型，水痘带状疱疹病毒，腮腺炎病毒，肠道病毒，人双埃可病毒及内部对照。
Ready to use lyo master mix (8-well strips each) for detection of herpes simplex virus 1, herpes simplex virus 2, varicella zoster virus, mumps virus, enterovirus, human parechovirus including internal control.

病毒性脑膜炎5联荧光PCR检测试剂盒

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【市场部】    杨永汉

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【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期2幢101-103室

五、实验步骤
1.菌液制备：
（1）实验前14—16小时，从冰箱保存的斜面菌种，挑少量菌于盛有5毫升*液体培养基的三角烧瓶中；每一个菌株接种一瓶，共接种4瓶，置37℃培养过夜。
（2）取出培养过夜的细菌，在W1177一瓶菌液中加入5毫升新鲜的*培养液，充分摇匀，等量分成2瓶；其余3瓶菌液分别用灭菌的5毫升吸管，各吸出2.5毫升菌液，然后再各加入2.5毫升新鲜的*培养液，充分摇匀，各菌于37℃继续培养3—5小时。
（3）自温箱取出三角烧瓶，分别倒入离心管，菌株W1177倒二支离心管，其余菌株各倒入一支离心管，离心沉淀，3，500转/分，离心10分钟。
（4）倒去上清液，打匀沉淀，加入无菌水，离心洗涤3次，再加无菌水到原体积。
2.杂交——混合培养：
（1）取12支灭菌试管，每支吸入3毫升经融化的半固体培养基，并保温在45℃（保温温度不宜高，北方气温低，可降低半固体中的琼脂量）。
（2）12支试管分成三个杂交组合，即W1177×K12pro；W1177×W1485；W1177×HfrC。每个组合各4支试管，其中2支对照，2支混合菌液。
（3）对照组试管各吸F+或Hfr供体菌菌液1毫升，其余按杂交组合各吸供体菌和受体菌菌液0.5毫升，充分混匀。
（4）将各试管中含菌的半固体倒在有Vogel培养基底层的平板上，摇匀待凝，放37℃培养，48小时后观察。
微生物具有容易变异的特性，因此，在保藏过程中，必须使微生物的代谢处于zui不活跃或相对静止的状态，才能在一定的时间内使其不发生变异而又保持生活能力。
低温、干燥和隔绝空气是使微生物代谢能力降低的重要因素，所以，菌种保藏方法虽多，但都是根据这三个因素而设计的。

Fifth, experimental steps
1. Bacterial liquid preparation:
(1) 14-16 hours prior to the experiment, pick up a small amount of bacteria in a beaker containing 5 ml of complete liquid medium from a bevelled strain in a refrigerator. Each strain was inoculated with a bottle of 4 inoculation at 37 ° C C*te overnight.
(2) Remove the bacteria cultured overnight, add 5 ml of fresh complete culture solution to a bottle of W1177, shake well, and divide into 2 bottles equally; the remaining 3 bottles of bacteria are sterilized 5 ml pipette, each Aspirate 2.5 ml of broth, and then add 2.5 ml of fresh complete broth, shake well, and each strain at 37 ° C for 3-5 hours.
(3) Remove the Erlenmeyer flask from the thermostat and pour into the centrifuge tube separay. The strain W1177 was poured into two centrifuge tubes. The remaining strains were each poured into a centrifuge tube and centrifuged at 3,500 rpm for 10 minutes.
(4) pour the supernatant, beat evenly, add sterile water, centrifuged and washed three times, plus sterile water to the original volume.
2. Hybridization - Mixed culture:
(1) Take 12 sterilized test tubes, each inhaled 3 ml of the thawed semi-solid medium, and incubated at 45 ° C (holding temperature should not be high, the northern temperature is low, can reduce the amount of semi-solid agar).
(2) Twelve tubes were divided into three hybrid combinations, namely W1177 × K12pro, W1177 × W1485 and W1177 × HfrC. Each combination of four test tubes, of which two control, two mixed bacteria solution.
(3) In the control group, each tube of F + or Hfr donor bacteria was drawn 1 ml, and the remaining 0.5 ml of the donor bacterium and the recipient bacteria were mixed by the hybridization, and the mixture was thoroughly mixed.
(4) Pour the semi-solid containing bacteria in each test tube on the plate with Vogel medium bottom, shake well and let stand and incubate at 37 ℃ for 48 hours.
Microbes have the characteristics of easy variation, therefore, in the preservation process, the metabolism of microorganisms must be in the most inactive or relatively static state, in a certain period of time so that it will not mutate while maintaining viability.
Cold, dry and isolated air is an important factor to reduce microbial metabolic capacity, so there are many ways to save bacteria, but are based on these three factors and design.