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 pneumoniae without cross-reaction with other biological genomes.

2. High sensitivity, the lowest detection limit is less than 10 genomes per reaction.

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 dUTP and UDG enzymes, which can reduce the contamination of residual DNA.

Mycoplasma pneumoniae is one of the smallest self-replicating organisms and is the human pathogen that causes mycoplasma pneumonia. The cells are elongated in shape, about 1-2 μm long and 0.1-0.2 μm wide, and cannot be detected under a light microscope; the length of the colonies is usually less than 100 μm and requires a stereomicroscope to observe. Mycoplasma pneumoniae parasitizes the human respiratory epithelium, adheres to respiratory epithelial cells by attaching organelles, and then fuses with host cells and enters the cells, evading detection by the host immune system, and adjusting the composition of the cell membrane to mimic the host cell membrane. M. pneumoniae is therefore prone to chronic or latent infection and, because its cell membrane composition is similar to human cells, may lead to autoimmune reactions in several organs and tissues.

Mycoplasma pneumoniae is distributed all over the world and can cause a variety of symptoms, including: pneumonia, Bronchitis, an upper respiratory tract disease, sometimes causes no symptoms and is often associated with otherBacteria or viruses are difficult to distinguish. Mycoplasma pneumoniae is frequently found in patients with respiratory illnesses and has also been linked to polyarthritis, stroke, infectious neuronitis, coronary artery disease, and more. Mycoplasma pneumoniae is an important causative agent of pneumonia and is reported to account for 10-30% of all acquired pneumonia cases in the general population and 25-71% in closed groups .

Mycoplasma pneumoniae grows slowly when cultured in vitro, sometimes taking several weeks, so In vitro culture assays are rarely used. Other possible detection methods include: Western blot, immunofluorescence staining, hemoadsorption test, tetrazolium reduction, metabolic inhibition test, serology test and polymerase chain reaction, etc. Due to its low cost and relatively short testing time, EIA serological analysis is the most commonly used method for detecting Mycoplasma pneumoniae. The disadvantage is that it requires the use of biopsied tissue, and this sampling method may increase the severity of the infection. Serological methods usually have low specificity and sensitivity and can only be used retrospectively. PCR is the fastest and most effective method to determine the presence of Mycoplasma pneumoniae. It has higher sensitivity and stronger specificity, and the detection time is short, and the results can be obtained in just a few hours. Compared with conventional PCR method, quantitative PCR method can not only accurately quantify, but also be more convenient to operate and less affected by environmental pollution.

This kit selects the cell adhesion gene as the target to specifically identify pneumonia Mycoplasma, verified by BLAST, has no cross-reactivity with other biological genomes. This kit detected 28 types of bacteria and 6 types of viruses with similar distribution ranges, and no non-specific signals were found. The DNA of 49 tissue samples was tested and 38 were found to be positive, which was consistent with the detection results of conventional PCR methods.

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 fromThe extremely thermophilic bacterium 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， 特异性检测肺炎支原体，与其他生物基因组没有交叉反应。

2， 灵敏度高，最低检测极限低于每反应10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dUTP和UDG酶，可降低残留DNA的污染。

 

肺炎支原体（Mycoplasma Pneumoniae）属于最小的自我复制型生物之一，是导致支原体肺炎的人类病原体。其细胞呈细长形状，长约1-2μm，宽约0.1-0.2μm，光学显微镜下无法检测；其菌落的长度通常小于100μm，需要立体显微镜来观察。肺炎支原体寄生于人类的呼吸道上皮，通过附着细胞器粘附于呼吸上皮细胞，然后与宿主细胞融合并进入细胞内，逃避宿主免疫系统的检测，并调节细胞膜的组成以模拟宿主细胞膜。因此肺炎支原体易于产生慢性或潜伏性感染，而且因其细胞膜组成与人细胞相似，可能导致几种器官和组织中的自身免疫反应。

肺炎支原体分布遍及全球，可引起多种症状，包括：肺炎、支气管炎、上呼吸道疾病，有时并不表现出症状，通常与其他细菌或病毒难以区分。肺炎支原体经常在患呼吸道疾病的病人中被发现，并且还与多发性关节炎、中风、传染性神经元炎、冠状动脉疾病等有关。肺炎支原体是肺炎的重要致病因子，据报道，在大众群体中占所有获得性肺炎病例的10-30%，在封闭性群体（如住宿的学生、军人）中则占25-71%。

肺炎支原体体外培养时生长缓慢，有时需数周的时间，因此很少使用体外培养法检测。其他可能的检测方法包括：免疫印迹，免疫荧光染色，血细胞吸附试验，四唑还原，代谢抑制试验，血清学试验和聚合酶链式反应（PCR）等。由于成本低，测试时间相对较短，EIA血清学分析是最常用的肺炎支原体检测方法，其缺点在于需要使用活组织，这种取样方式可能会加大感染的严重程度。血清学方法通常特异性和灵敏度都比较低，且只能做回顾性检测。PCR是确定肺炎支原体存在的最快速和最有效的方法，有更高的灵敏度和更强的特异性，且检测时间短，仅需几个小时即可获得结果。定量PCR法与常规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。

本试剂盒选择细胞黏附素基因作为靶点，特异性识别肺炎支原体，经BLAST验证，与其他生物基因组没有交叉反应。本试剂盒检测了分布范围类似的28种细菌和6种病毒，未发现非特异信号；对49个组织样品DNA进行检测，发现38个阳性，与常规PCR方法检测结果一致。

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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