呼吸道病原體21種多重檢試劑盒（PCR方法）

廣州健侖生物科技有限公司

準(zhǔn)備使用lyo master混合物（每個8孔帶）用于檢測：甲型流感，甲型流感（H1N1），乙型流感，冠狀病毒NL63,229E，OC43和HKU1，副流感1,2,3和4，人偏肺病毒 A和B，鼻病毒，呼吸道合胞病毒A和B，腺病毒，腸道病毒，小RNA病毒，博卡病毒，肺炎支原體包括內(nèi)部對照。
Ready to use lyo master mix (8-well strips each) for detection of: influenza A, influenza A (H1N1)swl, influenza B, coronaviruses NL63, 229E, OC43 and HKU1, parainfluenza 1, 2, 3 and 4, human metapneumovirus A and B, rhinovirus, respiratory syncytial viruses A and B, adenovirus, enterovirus, parechovirus, bocavirus, Mycoplasma pneumoniae including internal control.

呼吸道病原體21種多重檢試劑盒（PCR方法）

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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  】 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

（3）DAB變質(zhì)和顯色時間太長：DAB現(xiàn)用現(xiàn)配，如有沉渣應(yīng)進(jìn)行過濾后再用。配制好的DAB不應(yīng)存放時間太長，因為在沒有酶的情況下，*也會游離出氧原子與DAB產(chǎn)生反應(yīng)而降低DAB的效力，未用完的DAB存放在冰箱里幾天后再用這種似乎節(jié)約的辦法是不可取的。DAB的顯色在顯微鏡下監(jiān)控，達(dá)到理想的染色程度時立即終止反應(yīng)。不過當(dāng)染色片太多時或用染色機(jī)時，這樣做似乎不現(xiàn)實，但至少應(yīng)對一些新的或少用的抗體顯色時進(jìn)行監(jiān)控，避免顯色時間過長。
（4）組織變干：修復(fù)液溢出后未及時補(bǔ)充液體、染色切片太多、動作太慢、忘記滴液、滴液流失等都是造成組織變干的原因。解決的辦法是操作要認(rèn)真仔細(xì)，采用DAKO筆或PAPPen在組織周圍畫圈，可以有效的避免液體流失，也能提高操作速度。
（5）切片在緩沖液或修復(fù)液中浸泡時間太長（大于24小時）：原因上不清楚，但現(xiàn)象存在。有的實驗室喜歡前一天將切片脫蠟至修復(fù)，第二天加抗體進(jìn)行免疫組化染色，如果將裝有切片和修復(fù)液的容器放在4度冰箱過夜，對結(jié)果無明顯影響，如果放在室溫，特別是炎熱的夏天，會出現(xiàn)背景著色，因此，不可存放時間太長。
（6）一抗變質(zhì)、質(zhì)量差的多克隆抗體：注意抗體的有效期，過期的抗體要麼不顯色要麼背景著色。用新買的抗體時設(shè)立陽性對照和用使用過的抗體作比較。
2、切片邊緣著色
切片邊緣著色也是一種常見的現(xiàn)象，這種現(xiàn)象稱為邊緣效應(yīng)。產(chǎn)生的原因：（1）組織邊緣與玻片粘貼不牢，邊緣組織松脫漂浮在液體，每次清洗不易將組織下面試劑洗盡所致。解決辦法：制備優(yōu)質(zhì)的膠片（APES或多聚賴氨酸），切出盡量薄的組織切片，不厚于4微米，組織的前期處理應(yīng)規(guī)范，盡量避免選用壞死較多的組織。（2）切片上滴加的試劑未充分覆蓋組織，邊緣的試劑容易首先變干，濃度較中心組織高而致染色深。解決辦法：試劑要充分覆蓋組織，應(yīng)超出組織邊緣 2 mm。用DAKO筆畫圈時，為了避免油劑的影響，畫圈應(yīng)距組織邊緣3-4 mm。

(3) DAB deterioration and color development time is too long: DAB is best used now with sediment should be filtered before reuse DAB prepared should not be stored for too long, because in the absence of enzymes, peroxidation Hydrogen will also free oxygen atoms react with DAB to reduce the effectiveness of DAB, DUB stored in the refrigerator a few days and then use this seemingly economical approach is not desirable.DAB color is best in the microscope Under the control, to achieve the desired degree of staining immediay terminate the reaction. However, this may not be the case when there are too many dyed sheets or when using a dyeing machine, but at least some new or less used antibodies should be monitored during their color development to avoid excessive color development.
(4) tissue dry: repair fluid overflow after the liquid is not replenished in time, too many dyed slices, action is too slow, forgetting dripping, drip loss and so is the cause of tissue dry out The solution is to operate carefully, Use DAKO pen or PAPPen circle around the organization, can effectively avoid liquid loss, but also improve operating speed.
(5) Slides immersed in buffer or repair fluid for too long (more than 24 hours): reason is unclear, but the phenomenon exists Some laboratories like to deparaffinize the sections the day before to repair, and the next day the antibodies are immunized For histochemical staining, if the container containing sliced ??and reconstituted fluid is placed in a 4-degree refrigerator overnight, there is no noticeable effect on the result. If it is stored at room temperature, especially in the hot summer, background coloring occurs and therefore, it can not be stored for too long .
(6) polyclonal antibodies of the first anti-deterioration, poor quality: pay attention to the validity of the antibody, the expired antibodies either not color or the background color with the new antibody is best to set up a positive control and used antibodies for comparison.
2, slicing edge coloring
Slice edge coloring is also a common phenomenon, this phenomenon is called the edge effect of the reasons: (1) the edge of the tissue and the glass paste is not strong, the edge of tissue loose floating in the liquid, each cleaning is not easy to wash the tissue under the reagent Make the best solution: Prepare high-quality film (APES or polylysine), cut the tissue slices as thin as possible, not thicker than 4 microns, the organization's pre-treatment should be standardized, try to avoid using more necrotic tissue. (2) The reagent dropped on the slice is not enough to cover the tissue. The reagent on the edge is easy to dry at first, and the concentration is higher than the center tissue, resulting in deep staining. The specimen should be adequay covered with tissue and should be 2 mm beyond the edge of the tissue. With DAKO pen circle, in order to avoid the influence of oil, painted circle should be 3-4 mm from the edge of the organization.