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 Mycoplasma felis and Mycoplasma canis without interference from other genomes.

2. High sensitivity, and can detect at least 2 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.

 

Haemophilic Mycoplasma can infect Many mammals, including cats, dogs, pigs, cattle, sheep, etc., as well as humans. Mycoplasma haemofelis, also known as Mycoplasma felis and Eperythroblastoma felis, parasitizes on the surface of cat red blood cells. It is pleomorphic and can be spherical, ring-shaped or rod-shaped. It has a destructive effect on red blood cells and can cause infectious hemolytic anemia. Sometimes it has an acute attack, which can be fatal; sometimes it has a chronic attack, with symptoms such as lethargy, anorexia, pale mucous membranes, jaundice, fever, and splenomegaly, and may occur repeatedly. . The route of transmission is through blood, including bites from blood-sucking insects, bites, blood transfusions, etc.

There are currently three types of mycoplasma known to be parasitic in cat blood: Mycoplasma felis ,Candidatus Mycoplasma haemominutum and Candidatus Mycoplasma turicensis. Before the advent of PCR technology, these three mycoplasma species were collectively known as Haemobartonella felis. These bacteria adhere to and grow on the surface of red blood cells and sometimes 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. Mycoplasma felis is more pathogenic than the other two mycoplasmas. Double or triple infection often occurs with these three mycoplasmas. Mycoplasma felis is often the one with the smallest detected copy number, so a separate detection method is needed to eliminate interference from the other two mycoplasmas.

There are two types of mycoplasma that parasitize in dog blood: Mycoplasma canis and Candidatus Mycoplasma haematoparvum. Mycoplasma canis and Mycoplasma felis are very closely related, have similar morphologies, and their gene sequences are almost 100% identical. Under experimental conditions, cats can be infected with Mycoplasma canis, but dogs cannot be infected with Mycoplasma felis. Possible clinical manifestations of Mycoplasma canis include: asymptomatic, dehydration, mild anemia, tachycardia, jaundice, and death, but rarely causes obvious hemolysis unless splenectomy or immunosuppression occurs. Mycoplasma canis is the most common hemophilic mycoplasma in dogs and is extremely widely distributed, with a positive rate of approximately 0.5-40%.

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 Mycoplasma felis and Mycoplasma canis, and can be used with CandidatusMycoplasma haemobos has weak cross-reactivity. Candidatus Mycoplasma haemobos is a parasite in cattle blood and will not affect the detection of this kit. 1585 cat blood samples were tested and 45 positive results were obtained, 25 of which were co-infected with two other feline haemophilic mycoplasma species.

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 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， 特异性检测猫血支原体和犬血支原体，不受其他基因组干扰。

2， 灵敏度高，最低可检出反应液中的2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。

 

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。猫血支原体（Mycoplasma haemofelis），或称为猫嗜血支原体、猫附红细胞体，寄生于猫红细胞表面，具多形性，可呈现为球状、环状或杆状，对红细胞有破坏作用，可引起传染性溶血贫血症，有时呈急性发作，可致命；有时呈慢性发作，出现嗜睡、厌食、粘膜惨白及黄疸、发烧、脾脏肿大等症状，并可能出现反复发作。其传播途径为血液传播，包括：吸血昆虫叮咬、咬伤、输血等。

目前已知寄生于猫血中的支原体有三种：猫血支原体，Candidatus Mycoplasma haemominutum和Candidatus Mycoplasma turicensis。在PCR技术出现之前，这三种支原体被统称为猫血巴尔通氏体（Haemobartonella felis）。这些细菌粘附在红细胞表面生长，有时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。猫血支原体的致病能力强于另两种支原体。这三种支原体常发生双重或三重感染，猫血支原体往往是检出拷贝数最少的，因此需要单独的检测方法排除另两种支原体的干扰。

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。犬血支原体与猫血支原体亲缘关系非常近，形态也类似，基因序列几乎100%一致。在实验条件下，猫可以感染犬血支原体，而犬不能感染猫血支原体。犬血支原体可能的临床表现有：无症状、脱水、轻度贫血、心跳过速、黄疸、死亡，但是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。犬血支原体是犬类中最常见的嗜血支原体，分布极为广泛，阳性率大约在0.5-40%之间。

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，可以准确识别猫血支原体和犬血支原体，与Candidatus Mycoplasma haemobos有微弱的交叉反应。Candidatus Mycoplasma haemobos寄生于牛血中，不会对本试剂盒的检测产生影响。对1585个猫血样品进行检测，获得45个阳性，其中有25个为与另两种猫嗜血支原体共感染。

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。

 

 

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