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Probe-quantitative Real-time PCR Kit for Candidatus Mycoplasma haematoparvum
Probe-quantitative Real-time PCR Kit for Candidatus Mycoplasma haematoparvum
探针法Candidatus Mycoplasma haematoparvum实时定量PCR试剂盒
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Store in the dark at -20℃, transport at 2-8℃, valid for one year. Avoid repeated freezing and thawing.

 

 

Kit Features:

1. This kit can specifically detectCandidatus< /em> Mycoplasma haematoparvum, is not affected by microorganisms carried by dogs.

2. This kit has high sensitivity, and the lowest detection value is close to 1 genome.

3. The DNA polymerase used in this kit has the characteristics of strong resistance to inhibition and good thermal stability; it adopts hot start This method can inhibit non-specific amplification and reduce background fluorescence.

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

5, with dUTP and UDG enzymes, which 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 two known mycoplasmas that parasitize in dog blood: Mycoplasma haematoparvum and Candidatus Mycoplasma haematoparvum . Candidatus Mycoplasma haematoparvum was first discovered in a dog after splenectomy in 2004. It was subsequently found to be widely distributed in many regions around the world, with a positive rate ranging from 0-33.3%. Candidatus Mycoplasma haematoparvum is similar to Candidatus Mycoplasma haemominutum, with a diameter of 0.3 microns, no clustering phenomenon, and rarely causes obvious hemolysis unless splenectomy surgery or immunosuppression occurs.


Haemophilus Mycoplasma cannot be cultured in vitro, so it is difficult to develop protein detection methods. Commonly used detection methods include microscopy and PCR. The microscopy method has low sensitivity and large experimental errors. It cannot distinguish the types of mycoplasma and is easily interfered by other substances in the blood and artificial smear methods. Mycoplasma can easily fall off the surface of red blood cells, resulting in missed detection. The PCR method has obvious advantages in sensitivity and can distinguish mycoplasma species. 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 to design specific primers and The probe can accurately identify Candidatus Mycoplasma haematoparvum and is not affected by microorganisms carried by dogs. Testing of 184 canine blood samples yielded 7 positive results, 2 of which were co-infected with Mycoplasma canis.


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 extremothermophilic bacteria, 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 antibody is removed by heating, and the activity of the DNA polymerase is restored at this time, thus obtaining "thermal denaturation"."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, 本试剂盒可以特异性检测Candidatus Mycoplasma haematoparvum,不受犬类携带微生物的影响。

2, 本试剂盒灵敏度高,最低检出值接近1个基因组。

3, 本试剂盒使用的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, 本试剂盒带有阳性对照样品(组分C),可用于检验试剂盒有效性。

5, 带有dUTP和UDG酶,可降低残留DNA的污染。

 

嗜血支原体可以感染很多哺乳动物,包括猫、犬、猪、牛、羊等,也包括人类。目前已知寄生于犬血中的支原体有两种:犬血支原体(Mycoplasma haemocanis,旧称Haemobartonella canis)和Candidatus Mycoplasma haematoparvum。Candidatus Mycoplasma haematoparvum于2004年在一只行脾切除手术后的犬中首次被发现,随后发现其在世界多个地区广泛分布,阳性率在0-33.3%不等。Candidatus Mycoplasma haematoparvum与Candidatus Mycoplasma haemominutum较为相似,直径为0.3微米,无成串相连现象,很少引起明显的溶血,除非是做了脾切除手术或者发生了免疫抑制。

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 haematoparvum,不受犬类携带微生物的影响。对184个犬血样品的检测获得7个阳性,其中2个为与犬血支原体共感染。

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。

 

 

 

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