呼吸道病原体21种多重检试剂盒（PCR方法）

广州健仑生物科技有限公司

准备使用lyo master混合物（每个8孔带）用于检测：甲型流感，甲型流感（H1N1），乙型流感，冠状病毒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，肠道病毒，小RNA病毒，博卡病毒，肺炎支原体包括内部对照。
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.

呼吸道病原体21种多重检试剂盒（PCR方法）

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【公司名称】 广州健仑生物科技有限公司
【市场部】    杨永汉

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【腾讯  】 2042552662
【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

（3）DAB变质和显色时间太长：DAB现用现配，如有沉渣应进行过滤后再用。配制好的DAB不应存放时间太长，因为在没有酶的情况下，过氧化氢也会游离出氧原子与DAB产生反应而降低DAB的效力，未用完的DAB存放在冰箱里几天后再用这种似乎节约的办法是不可取的。DAB的显色在显微镜下监控，达到理想的染色程度时立即终止反应。不过当染色片太多时或用染色机时，这样做似乎不现实，但至少应对一些新的或少用的抗体显色时进行监控，避免显色时间过长。
（4）组织变干：修复液溢出后未及时补充液体、染色切片太多、动作太慢、忘记滴液、滴液流失等都是造成组织变干的原因。解决的办法是操作要认真仔细，采用DAKO笔或PAPPen在组织周围画圈，可以有效的避免液体流失，也能提高操作速度。
（5）切片在缓冲液或修复液中浸泡时间太长（大于24小时）：原因上不清楚，但现象存在。有的实验室喜欢前一天将切片脱蜡至修复，第二天加抗体进行免疫组化染色，如果将装有切片和修复液的容器放在4度冰箱过夜，对结果无明显影响，如果放在室温，特别是炎热的夏天，会出现背景着色，因此，不可存放时间太长。
（6）一抗变质、质量差的多克隆抗体：注意抗体的有效期，过期的抗体要麽不显色要麽背景着色。用新买的抗体时设立阳性对照和用使用过的抗体作比较。
2、切片边缘着色
切片边缘着色也是一种常见的现象，这种现象称为边缘效应。产生的原因：（1）组织边缘与玻片粘贴不牢，边缘组织松脱漂浮在液体，每次清洗不易将组织下面试剂洗尽所致。解决办法：制备优质的胶片（APES或多聚赖氨酸），切出尽量薄的组织切片，不厚于4微米，组织的前期处理应规范，尽量避免选用坏死较多的组织。（2）切片上滴加的试剂未充分覆盖组织，边缘的试剂容易首先变干，浓度较中心组织高而致染色深。解决办法：试剂要充分覆盖组织，应超出组织边缘 2 mm。用DAKO笔画圈时，为了避免油剂的影响，画圈应距组织边缘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.