Details
Store in the dark at -20℃, transport at 2-8℃, valid for one year. Avoid repeated freezing and thawing.
Kit Features:
1. Specific detection of Candidatus Mycoplasma turicensis, with no cross-reactivity with other biological genomes.
2. 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.
3. With positive control sample, which can be used to test the effectiveness of the kit.
4. With dUTP and UDG enzymes, it can reduce the contamination of residual DNA.
Haemophilic Mycoplasma can infect many mammals, including cats, dogs, pigs, cattle, sheep, etc., as well as humans. There are currently three known mycoplasmas that parasitize in cat blood: Mycoplasma haemofelis), Candidatus Mycoplasma haemominutum and Candidatus Mycoplasma turicensis. These bacteria adhere to and grow on the surface of red blood cells and can also cause some diseases to the host, especially when the cat's immunity is weakened. It has been reported that about 14% of anemic cats are positive for blood mycoplasma. Candidatus Mycoplasma turicensis parasitizes on the surface of cat red blood cells. It has moderate pathogenicity among the above three mycoplasmas, and is often co-infected with the other two mycoplasmas. It can cause mild to acute hemolytic anemia and is related to cat immunity. Clinical symptoms worsen when the defective virus is co-infected or after treatment with cortisone drugs.
Haemophilus Mycoplasma cannot be cultured in vitro, so it is difficult to develop protein detection methods. Commonly used detection methods include microscopy and PCR. Microscopy The sensitivity of the method is low, the experimental error is large, the type of mycoplasma cannot be distinguished, and it is easily interfered by other substances in the blood and artificial smear methods, and the mycoplasma is easy to fall off from the surface of red blood cells, resulting in missed detection. The PCR method has obvious advantages in sensitivity. , and can distinguish the types of mycoplasma. 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 is based on the conserved region of the 16S rRNA gene and designs specific primers and probes that can accurately identify Candidatus Mycoplasma turicensis, has no cross-reactivity with other biological genomes. Among 236 cat blood samples, 14 positive results were obtained, of which 2 were infected with Candidatus Mycoplasma turicensis alone, 2 were co-infected with Mycoplasma felis, and 3 were infected withCandidatus Mycoplasma haemominutum co-infection, and 7 three haemophilic mycoplasma co-infections.
This kit contains hot-start DNA polymerase, dNTP, primers, probes, positive controls, ROX dye, and UDG to prevent residual DNA contamination , uracil-DNA-glycosylase), users only need to prepare DNA samples to use.
The DNA polymerase used in this kit is derived from the extremely thermophilic bacterium . After modification, it has stronger resistance to inhibition and thermal stability than ordinary Taq enzymes. property. 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 removed by heat, and DNA polymerase activity is restored at this time, thus obtaining the "hot start" function. Hot start priming reduces residual DNA contamination and improves PCR amplification efficiency, sensitivity and target fragment yield.
UDG and dUTP are used to eliminate residual contamination in PCR amplification. dUTP ensures that amplified DNA contains uracil. UDG removes uracil residues from single- or double-stranded DNA, thereby preventing uracil-containing DNA from being used as a template for further amplification. UDG is inactivated by high temperature during normal PCR cycles and has no effect on the PCR reaction.
-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， 特异性检测Candidatus Mycoplasma turicensis，与其他生物基因组没有交叉反应。
2， 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。
3， 带有阳性对照样品（组分C），可用于检验试剂盒有效性。
4， 带有dUTP和UDG酶，可降低残留DNA的污染。
嗜血支原体（Hemoplasma）可以感染很多哺乳动物，包括猫、犬、猪、牛、羊等，也包括人类。目前已知寄生于猫血中的支原体有三种：猫血支原体（Mycoplasma haemofelis），Candidatus Mycoplasma haemominutum和Candidatus Mycoplasma turicensis。这些细菌粘附在红细胞表面生长，也会给宿主造成一些疾病，尤其是在猫免疫力下降的时候。有报道约14%的贫血猫呈血液支原体阳性。Candidatus Mycoplasma turicensis寄生于猫红细胞表面，在上述三种支原体中致病能力中等，且常与另两种支原体合并感染，可引起温和到急性的溶血性贫血，与猫免疫缺陷病毒合并感染时，或者可的松类药物处理后，临床症状加重。
嗜血支原体无法在体外培养，因此难以开发蛋白类检测方法。常用的检测方法包括显微镜镜检法和PCR法。镜检法灵敏度低，实验误差大，无法区分支原体种类，易受血液中其他物质以及人工涂片方法的干扰，且支原体易从红细胞表面脱落，造成漏检。而PCR法在灵敏度上则有明显的优势，且可以区分支原体种类。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。
本试剂盒基于16S rRNA基因的保守区域，设计特异性引物和探针，可以准确识别Candidatus Mycoplasma turicensis，与其他生物基因组没有交叉反应。在236份猫血样品中，检测获得14个阳性，其中2个为Candidatus Mycoplasma turicensis单独感染，2个为与猫血支原体共感染，3个为与Candidatus Mycoplasma haemominutum共感染，以及7个三种嗜血支原体共感染。
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），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。
UDG和dUTP用于消除PCR扩增中的残余污染。dUTP可以确保扩增后的DNA中均含有尿嘧啶。UDG可从单链或双链DNA上去除尿嘧啶残基，从而阻止含有尿嘧啶的DNA作为模板继续扩增。UDG在正常的PCR循环过程中被高温灭活，对PCR反应没有影响。
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