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Probe-quantitative Real-time PCR Kit for Candidatus Mycoplasma haemolamae
Probe-quantitative Real-time PCR Kit for Candidatus Mycoplasma haemolamae
探针法Candidatus Mycoplasma haemolamae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. Specific detection of Candidatus Mycoplasma haemolamae, no cross-reaction with other biological genomes.

2. High sensitivity, the detection rate is 90% when there is only one genome in the reaction solution. The linear range spans 8 orders of magnitude.

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 is a type of bacteria that lives on the surface of red blood cells. It has no cell wall and can infect many mammals, including cats, dogs, pigs, cattle, sheep, etc. Including humans. Haemophilus Mycoplasma was once named Hemobartonella and Eperythrozoon . Based on 16S rRNA gene sequence analysis, it was reclassified as a mycoplasma. . Candidatus Mycoplasma haemolamae is parasitic in the blood of camels in South America and was first reported in 1990. Affected by the condition of the host, the clinical manifestations after infection are diverse, which can be asymptomatic, or chronic weight loss, depression, decreased fecundity, lethargy, acute failure, and in rare cases death. Appropriate medication can improve animal healthbed conditions, reducing the bacterial load, but it may not be completely eliminated and eventually asymptomatic carriers will form

Candidatus There is no suitable in vitro culture for Mycoplasma haemolamae method, so it is difficult to develop protein detection methods. The detection method generally uses the Giemsa-stained blood smear method. The detection sensitivity and specificity of the blood smear method are relatively poor, and it is easily interfered by artificial preparation methods and other components in the blood, and 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 designs specific primers and probes based on the 16S rRNA gene. Accurately identify Candidatus Mycoplasma haemolamae without cross-reaction with other biological genomes. After testing, this kit has no cross-reactivity with 9 types of haemophilic mycoplasma from cats, dogs, cattle, pigs and mice, as well as 4 types of non-haemophilic mycoplasma. This kit can be used to specifically detect Candidatus Mycoplasma haemolamae.


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. DNA polymerase activity is inhibited at room temperature by binding to specific monoclonal antibodies. 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Candidatus Mycoplasma haemolamae,与其他生物基因组无交叉反应。

2, 灵敏度高,反应液中仅1个基因组时检出率为90%。线性范围跨越8个数量级。

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(Hemotropic Mycoplasma,或hemoplasma)是一类寄生于红细胞表面的细菌,无细胞壁,可以感染很多哺乳动物,包括猫、犬、猪、牛、羊等,也包括人类。嗜血支原体曾被命名为血巴通氏体(Hemobartonella)和附红细胞体(Eperythrozoon),根据16S rRNA基因序列分析,被重新归类为支原体。Candidatus Mycoplasma haemolamae寄生于南美骆驼血液中,最早报道于1990年。受宿主状况影响,其感染后临床表现多样,可表现为无症状,或慢性体重下降、抑郁、繁殖力下降、昏睡、急性衰竭,极少数情况下出现死亡。合适用药可改善动物临床状况,降低细菌载量,但是可能无法彻底清除而最后形成无症状带菌者。

Candidatus Mycoplasma haemolamae尚无合适的体外培养方法,因此难以开发蛋白类检测方法。检测方法一般采用吉姆萨染色的血涂片法。血涂片法的检测灵敏度和特异性均比较差,容易受到人工制片方法和血液里其他成分(如:Howell–Jolly体)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基因设计特异性引物和探针,可以准确识别Candidatus Mycoplasma haemolamae,与其他生物基因组无交叉反应。经检测,本试剂盒与猫、犬、牛、猪和鼠的9种嗜血支原体,以及4种非嗜血支原体,均无交叉反应。本试剂盒可用于特异性检测Candidatus Mycoplasma haemolamae。

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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