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 and differentiation of Ureaplasma parvum and Ureaplasma urealyticum, and can identify human Ureaplasma All fourteen serotypes showed 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.
Ureaplasma is a type of mycoplasma with the ability to decompose urea. It is spherical or club-shaped, Gram-positive, and its clones are omelette-shaped, with a cell diameter of 0.1 to 1.0 μm. It was first isolated from the human urogenital tract in 1954 and named Ureaplasma urealyticum in 1974. Human There are currently 14 known serotypes in China. After years of debate, in 2002, based on genome size, 16S rRNA gene sequence, 16S-23S rRNA gene spacer sequence, enzyme polymorphism, DNA-DNA hybridization, differentiation response to manganese, multi-band antigen genes, etc., It is distinguished into two organisms: Ureaplasma urealyticum and Ureaplasma parvum . The differentiated Ureaplasma urealyticum contains ten serotypes: 2, 4, 5, 7, 8, 9, 10, 11, 12 and 13. Ureaplasma parvum contains four serotypes: 1. 3, 6 and 14. There is currently no consistent conclusion on the impact of different serotypes on human diseases. These two species of Ureaplasma are healthyCommon urogenital tract commensal bacteria in Kang people can exist alone or together, and are sometimes associated with some diseases, such as: urethritis, postpartum endometritis, chorioamnionitis, spontaneous abortion, premature birth, low birth weight, Pneumonia, bacteremia, meningitis and chronic lung disease of premature infants, etc. It is unclear under what circumstances these Ureaplasmas cause disease, and people with intact humoral immune systems are generally unaffected.
The in vitro culture of Ureaplasma is very difficult, time-consuming and less sensitive. Low. The typing of Ureaplasma relies on monoclonal or polyclonal antibodies against whole cells or purified antigens. Experimental methods include: growth inhibition experiment, metabolic inhibition experiment, immunofluorescence, immunoperoxidase, ELISA, western blot, etc. These methods are time-consuming and labor-intensive, the results are often difficult to reproduce, there are many cross-reactions, and it is often difficult to distinguish between samples containing multiple serotypes. The PCR method used to detect Ureaplasma has higher sensitivity and better specificity, is not affected by contamination from other microorganisms, and has a short detection time, 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 characteristic urease gene of Ureaplasma as the target , compare the sequences of 14 serotypes of Ureaplasma, design primers and probes, and specifically identify and distinguish Ureaplasma urealyticum and Ureaplasma parvum. This kit can accurately distinguish the biological classification of 14 serotypes of Ureaplasma. It has been verified by BLAST that there is no cross-reaction with other biological genomes. 42 isolated and cultured samples were tested, and 37 Ureaplasma parvum positives, 4 Ureaplasma urealyticum positives, and one double positive were obtained, which were completely consistent with the sequencing results. 31 clinical swab samples were tested, and 11 were positive for Ureaplasma parvum and 6 were positive for Ureaplasma urealyticum, which was consistent with the conventional PCR method. The positive rate was higher than the in vitro culture method, showing that the kit has high sensitivity. in vitro culture method.
-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， 特异性检测并区分微小脲原体和解脲脲原体，可识别人类脲原体的全部十四种血清型，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。
脲原体（Ureaplasma）是一类具有尿素分解能力的支原体（Mycoplasma），呈球状或球杆状，革兰氏阳性，克隆呈煎蛋状，细胞直径在0.1到1.0 μm 之间，于1954年首次从人类泌尿生殖道分离出来，1974年被命名为解脲脲原体（Ureaplasma urealyticum，也称为溶脲脲原体），人类中目前已知有14种血清型。经过多年的争论，在2002年，根据基因组大小、16S rRNA基因序列、16S-23S rRNA基因间隔区序列、酶的多态性、DNA-DNA杂交、对锰的分化反应、多带抗原基因等，将其区分为两种生物：解脲脲原体和微小脲原体（Ureaplasma parvum，也称为细小脲原体）。区分后的解脲脲原体包含的血清型有十种：2、4、5、7、8、9、10、11、12和13，微小脲原体包含的血清型有四种：1、3、6和14。不同血清型对人体疾病的影响目前尚没有一致结论。这两种脲原体是健康人常见的泌尿生殖道共生菌，可以单独或共同存在，有时也会与一些疾病相关联，如：尿道炎、产后子宫内膜炎、绒毛膜羊膜炎、自然流产、早产、低出生体重、肺炎、菌血症、脑脊膜炎和早产儿慢性肺病等。目前尚不清楚这些脲原体会在什么情况下引起疾病，体液免疫系统健全的人一般不会受其影响。
脲原体的体外培养非常困难，耗时较长，灵敏度较低。对脲原体的分型是依靠针对全细胞或纯化抗原的单克隆或多克隆抗体，实验方法包括：生长抑制实验、代谢抑制实验、免疫荧光、免疫过氧化物酶、ELISA、免疫印迹等，这些方法都比较耗时费力，结果常常难以重复，有很多交叉反应，在含有多个血清型的样本上常难以区分。而PCR法用于检测脲原体，则有更高的灵敏度和更好的特异性，不受其他微生物污染的影响，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。
本试剂盒选择脲原体特征性的尿素酶基因作为靶点，将脲原体14种血清型的序列进行比对，设计引物和探针，特异性识别并区分解脲脲原体和微小脲原体。使用本试剂盒可以准确区分脲原体14种血清型的生物分类，经BLAST验证，与其他生物基因组没有交叉反应。对42个分离培养样本进行检测，得到37个微小脲原体阳性，4个解脲脲原体阳性，以及一个双重阳性，与测序结果完全一致。对31个临床拭子样本进行检测，获得11个微小脲原体阳性，6个解脲脲原体阳性，与常规PCR法检测结果一致，阳性率高于体外培养法，显示本试剂盒灵敏度高于体外培养法。
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