Store in the dark at -20℃, transport at 2-8℃, valid for one year. Avoid repeated freezing and thawing.

 

 

Kit Features:

1. It can specifically detect Mycoplasma conjunctiva without cross-reaction with other organisms.

2. High sensitivity, the detection limit is 4 genomes in the reaction solution.

3. The DNA polymerase used has the characteristics of strong anti-inhibition ability and good thermal stability; using hot start method, it can Inhibit non-specific amplification and reduce background fluorescence.

4, with positive control sample, which can be used to test the effectiveness of the kit.

5. With UDG enzyme and dUTP, it can reduce the contamination of residual DNA.

Mycoplasma conjunctiva is the main pathogen of infectious keratoconjunctivitis in ruminants such as sheep, goats, and antelopes, and is widely distributed in domestic ruminants around the world. IKC is a common infectious eye disease known as pinkeye of small ruminants that can be transmitted between domestic and wild animals and is characterized by inflammation of the conjunctiva and cornea. In advanced stages, the cornea becomes opaque or even perforated, blinding wild animals that may fall off cliffs or die of starvation. One study showed that the positivity rate for Mycoplasma conjunctiva antibodies in adult sheep was 53%. Mycoplasmal IKC can be transmitted between different species, possibly through physical contact or vector insects such as flies. The prevalence of M. conjunctivae is currently unknown in most countries, which may be due to a lack of technology to detect M. conjunctivae.


The classic diagnostic method for conjunctival Mycoplasma infection is in vitro culture and subsequent analysis of the mycoplasma colonies Immunological identification. butHowever, this method is very cumbersome, takes a long time, and requires specialized technology. The use of PCR detection has the characteristics of high sensitivity and high specificity, and the detection time is short, and the results can be obtained in just a few hours. Compared with the ordinary PCR method, the quantitative PCR method can not only accurately quantify, but also be more convenient to operate and less affected by environmental pollution.


This kit selects a membrane protein gene in the genome of Mycoplasma conjunctiva for PCR Identification: The primers were verified by BLAST to specifically target Mycoplasma conjunctivalis and had no cross-reactivity with the genomes of other organisms. By detecting closely related mycoplasma and other microorganisms that parasitize sheep, this kit is specific for mycoplasma conjunctiva and has no false positive results. Swab samples from 182 goats, sheep or antelopes were tested. The detection sensitivity of this kit was significantly better than the nested PCR method of parallel detection.


This kit contains hot-start DNA polymerase, dNTP, primers, probes, positive controls, ROX dye, and UDG to prevent residual DNA contamination. Users only need to prepare DNA samples and use them.


The DNA polymerase used in this kit is derived from Thermus thermophilus HB-B), which has been modified to have stronger resistance to inhibition and thermal stability than ordinary Taq enzymes. By binding specific monoclonal antibodies, DNA polymerase activity is inhibited at room temperature. During the thermal denaturation phase of the PCR cycle, the antibodies are heated and removed, and the DNA polymerase activity is restored at this time, thus obtaining the "hot start" function. Hot start can reduce the contamination of residual DNA and improve PCR amplification efficiency, sensitivity and target fragment yield.

 

 

-20℃避光保存，2-8℃运输，有效期一年。避免反复冻融。

 

 

试剂盒特点：

1， 可以特异性检测结膜支原体，与其他生物无交叉反应。

2， 灵敏度高，检测极限为反应液中4个基因组。

3， 使用的DNA聚合酶具有抗抑制能力强和热稳定性好的特点；采用热启动方式，可抑制非特异性扩增，降低背景荧光。

4， 带有阳性对照样品（组分C），可用于检验试剂盒有效性。

5， 带有UDG酶和dUTP，可降低残留DNA的污染。

 

结膜支原体（Mycoplasma Conjunctivae）是绵羊、山羊、羚羊等反刍动物的传染性角膜结膜炎（IKC）的主要病原体，在全球广泛分布于家养的反刍类动物中。IKC是一种常见的传染性眼病，称为小反刍动物的红眼病，可在家养和野生动物之间传播，其特点是结膜和角膜的炎症。在晚期阶段，角膜变得不透明甚至穿孔，致盲的野生动物可能从悬崖坠落或死于饥饿。一项研究表明，成年绵羊中结膜支原体抗体的阳性率为53％。支原体性IKC可以在不同物种之间传播，传播途径可能是物理接触或苍蝇等媒介昆虫。目前在大多数国家尚不清楚结膜支原体的流行程度，这可能是由于缺乏检测结膜支原体的技术。

结膜支原体感染的经典诊断方法是体外培养和后续对支原体菌落的免疫学鉴定。但是这个方法非常繁琐，需要较长的时间，且需要专业化的技术。而使用PCR法检测则具有高灵敏度和高特异性的特点，且检测时间短，仅需几个小时即可获得结果。定量PCR法与普通PCR法相比，不仅可以精确定量，而且操作更为方便，更少受环境污染的影响。

本试剂盒选取了结膜支原体基因组内的一个膜蛋白基因进行PCR鉴定，引物经BLAST验证为特异性靶向结膜支原体，与其他生物的基因组无交叉反应。通过检测亲缘关系较近的支原体和寄生于羊的其他微生物，本试剂盒对结膜支原体具有特异性，未见假阳性结果。对来自182只山羊、绵羊或羚羊的拭子样本进行检测，本试剂盒检测灵敏度明显优于平行检测的巢式PCR法。

本试剂盒包含热启动DNA聚合酶、dNTP（以dUTP代替dTTP）、引物、探针、阳性对照品、ROX染料，和用以防止残余DNA污染的UDG（Uracil- DNA Glycosylase，尿嘧啶-DNA-糖基酶），用户只需准备好DNA样品即可使用。

本试剂盒采用的DNA聚合酶源自极端嗜热菌（Thermus thermophilus HB-B），经改造后较普通的Taq酶具有更强的抗抑制能力和热稳定性。通过结合特异性单克隆抗体，在室温下DNA聚合酶活性被抑制。在PCR循环的热变性阶段，抗体受热被去除，此时DNA聚合酶活性恢复，因此获得“热启动”功能。热启动可减少残余DNA的污染，提高PCR扩增效率、灵敏度和目的片段产量。

 

 

 

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