病毒性腦膜炎5聯(lián)熒光PCR檢測(cè)試劑盒

廣州健侖生物科技有限公司

準(zhǔn)備使用lyo master混合物（每個(gè)8孔條）檢測(cè)單純皰疹病毒1型，單純皰疹病毒2型，水痘帶狀皰疹病毒，腮腺炎病毒，腸道病毒，人雙埃可病毒及內(nèi)部對(duì)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.

病毒性腦膜炎5聯(lián)熒光PCR檢測(cè)試劑盒

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

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【騰訊  】 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(hào)二期2幢101-103室

五、實(shí)驗(yàn)步驟
1.菌液制備：
（1）實(shí)驗(yàn)前14—16小時(shí)，從冰箱保存的斜面菌種，挑少量菌于盛有5毫升*液體培養(yǎng)基的三角燒瓶中；每一個(gè)菌株接種一瓶，共接種4瓶，置37℃培養(yǎng)過夜。
（2）取出培養(yǎng)過夜的細(xì)菌，在W1177一瓶菌液中加入5毫升新鮮的*培養(yǎng)液，充分搖勻，等量分成2瓶；其余3瓶菌液分別用滅菌的5毫升吸管，各吸出2.5毫升菌液，然后再各加入2.5毫升新鮮的*培養(yǎng)液，充分搖勻，各菌于37℃繼續(xù)培養(yǎng)3—5小時(shí)。
（3）自溫箱取出三角燒瓶，分別倒入離心管，菌株W1177倒二支離心管，其余菌株各倒入一支離心管，離心沉淀，3，500轉(zhuǎn)/分，離心10分鐘。
（4）倒去上清液，打勻沉淀，加入無菌水，離心洗滌3次，再加無菌水到原體積。
2.雜交——混合培養(yǎng)：
（1）取12支滅菌試管，每支吸入3毫升經(jīng)融化的半固體培養(yǎng)基，并保溫在45℃（保溫溫度不宜高，北方氣溫低，可降低半固體中的瓊脂量）。
（2）12支試管分成三個(gè)雜交組合，即W1177×K12pro；W1177×W1485；W1177×HfrC。每個(gè)組合各4支試管，其中2支對(duì)照，2支混合菌液。
（3）對(duì)照組試管各吸F+或Hfr供體菌菌液1毫升，其余按雜交組合各吸供體菌和受體菌菌液0.5毫升，充分混勻。
（4）將各試管中含菌的半固體倒在有Vogel培養(yǎng)基底層的平板上，搖勻待凝，放37℃培養(yǎng)，48小時(shí)后觀察。
微生物具有容易變異的特性，因此，在保藏過程中，必須使微生物的代謝處于zui不活躍或相對(duì)靜止的狀態(tài)，才能在一定的時(shí)間內(nèi)使其不發(fā)生變異而又保持生活能力。
低溫、干燥和隔絕空氣是使微生物代謝能力降低的重要因素，所以，菌種保藏方法雖多，但都是根據(jù)這三個(gè)因素而設(shè)計(jì)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.