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Probe-quantitative Real-time PCR Kit for Chlamydia psittaci cluster
Probe-quantitative Real-time PCR Kit for Chlamydia psittaci cluster
探针法鹦鹉热衣原体族实时定量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 Waddlia chondrophila, no cross-reaction with other biological genomes.

2, High sensitivity, the lowest detection limit is about 80 copies/reaction.

3. DNA polymerase adopts hot start method. It can inhibit non-specific amplification and reduce background fluorescence.

4, with positive control sample, can be used Check the effectiveness of the kit.

5, with UDG enzyme and dUTP, can Reduce residual DNA contamination.

 

Chlamydia psittaci, also known as Chlamydophila psittaci/Chlamydia psittaci, Chlamydia psittaci, is a strictly intracellular parasitic prokaryotic organism belonging to the family Chlamydiaceae em>). Chlamydia psittaci is the earliest chlamydia discovered. It has a wide range of hosts, not only a variety of birds including parrots, but also many mammals; unique subtypes have been formed in many hosts, including: A–F, M56 and WC, each subtype has strict host specificity: the host of subtype A is parrots, the host of subtype B is pigeon, the host of subtype C is duck and goose, the host of subtype D is turkey, and the host of subtype E is turkey. Type 1 hosts are pigeons, ducks and other birds, F subtype hosts are parrots, WC subtype hosts are cattle, and M56 subtype hosts are rodents. In addition, there are some subtypes that are divided into independent species based on the differences in 16S and 23S rRNA sequences, such as: Chlamydia abortus, Chlamydia felis, guinea pig chlamydia and animal chlamydia, etc. Some of these chlamydiae can infect humans: Chlamydia psittaci can cause flu-like symptoms when infecting humans, sometimes developing into severe pneumonia, endocarditis or encephalitis; Chlamydia abortus can cause miscarriage and respiratory disorders in humans; Chlamydia feline can cause Conjunctivitis in humans; Chlamydia guinea pigs may cause pneumonia in humans. There are no reports of animal chlamydia infecting humans.

 

Chlamydia psittaci can be cultured and serologically method for detection. Chlamydia psittaci is easy to grow in chicken embryo yolk sac, HeLa cells, and monkey kidney cell cultures. However, because it is time-consuming and laborious and requires a high level of laboratory safety, the culture method is rarely used. Some serological methods, such as complement fixation test, indirect hemagglutination test technology, ELISA, and microimmunofluorescence method, are often used to detect Chlamydia psittaci. The disadvantage is that cross-reaction may occur and the sensitivity is low, making it unsuitable for early infection. Generally it can only be used for retrospective diagnosis. The PCR method is an in vitro enzymatic synthesis method of specific DNA fragments. It is more suitable for detecting Chlamydia psittaci. It has high sensitivity, strong specificity, and fast detection speed, which can be completed in only two or three hours. Compared with ordinary PCR method, quantitative PCR method is more convenient to operate and less affected by environmental pollution.

 

This kit uses probe method quantitative PCR to design primers based on the conserved sequences of extracellular membrane protein genes to specifically identify Chlamydia psittaci and Chlamydia abortus. , Chlamydia felis, Chlamydia guinea pig, Chlamydia buteonis and Chlamydia ibidis; these chlamydiae are all closely related to Chlamydia psittaci or were once classified as Chlamydia psittaci, and are related to Chlamydia psittaci Chlamydia trachomatis is distantly related. Verified by BLAST, this kit has no cross-reactivity with the genomes of other organisms. Use this kit to detect different chlamydia, as well as those that are closely related or relatively distant. No non-specific signals were found for 22 common bacteria. Chlamydia trachomatis, Chlamydia pneumoniae and veterinary Chlamydia were unable to produce specific signals; for 18 clinical samples Swabs and bronchial wash fluid) were tested and 8 positive results were obtained, which was more sensitive than the ELISA method and consistent with the clinical diagnosis results; 9 nasal swab samples originating from pigeons were tested and 3 positive results were obtained . This kit is suitable for the detection of Chlamydia psittaci family.

 

 

 

-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,   特异性检测鹦鹉热衣原体,与其他生物基因组无交叉反应。

2,   灵敏度高,最低检测极限约为80个拷贝/反应。

3,   DNA聚合酶采用热启动方式,可抑制非特异性扩增,降低背景荧光。

4,   带有阳性对照样品,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。

 

鹦鹉热衣原体(Chlamydia psittaci),或称鹦鹉热嗜衣原体/鹦鹉热亲衣原体(Chlamydophila psittaci)、鹦鹉衣原体,是一种严格细胞内寄生的原核生物,属于衣原体科(Chlamydiaceae)。鹦鹉热衣原体是最早发现的衣原体,宿主非常广泛,不仅有包括鹦鹉在内的多种鸟类,还有许多哺乳动物;在许多宿主中已形成独特的亚型,包括:A–F, M56和WC,每种亚型都具有严格的宿主特异性:A亚型宿主为鹦鹉科鸟类,B亚型宿主为鸽,C亚型宿主为鸭和鹅,D亚型宿主为火鸡,E亚型宿主为鸽、鸭及其他鸟类,F亚型宿主为鹦鹉,WC亚型宿主为牛,M56亚型宿主则为啮齿类动物。另外还有一部分亚型,根据16S和23S rRNA序列的差异,被划分为独立物种,如:流产衣原体(Chlamydia abortus)、猫衣原体(Chlamydia felis)、豚鼠衣原体(Chlamydia caviae)和兽类衣原体(Chlamydia pecorum)等。这些衣原体有些可以感染人类:鹦鹉热衣原体感染人类后可引起类似流感的症状,有时会发展为严重肺炎、心内膜炎或脑炎;流产衣原体可引起人类流产和呼吸功能紊乱;猫衣原体可引起人类的结膜炎;豚鼠衣原体可能引起人类肺炎。兽类衣原体尚无感染人类的报道。

鹦鹉热衣原体可使用培养法和血清学方法进行检测。鹦鹉热衣原体在鸡胚卵黄囊及HeLa细胞、猴肾细胞培养中易于生长,但由于耗时费力,且需要实验室安全等级较高,因此培养法很少使用。一些血清学方法,如:补体结合试验、间接血凝试验技术、ELISA、微量免疫荧光法,常用于检测鹦鹉热衣原体,其缺点在于可能产生交叉反应,且灵敏度较低,不适用于早期感染,一般只能用作回顾性诊断。PCR法是一种体外酶促合成特异性DNA 片段的方法,更适合检测鹦鹉热衣原体,其灵敏度高,且特异性强,检测速度快,只需两三个小时即可完成。定量PCR法与普通PCR法相比,操作更为方便,更少受环境污染的影响。

本试剂盒采用探针法定量PCR,针对胞膜外蛋白基因的保守序列设计引物,特异性识别鹦鹉热衣原体、流产衣原体、猫衣原体、豚鼠衣原体、Chlamydia buteonisChlamydia ibidis;这几种衣原体均与鹦鹉热衣原体亲缘关系很近或曾归属于鹦鹉热衣原体,而与沙眼衣原体(Chlamydia trachomatis)亲缘关系较远。经BLAST验证,本试剂盒与其他生物基因组没有交叉反应。使用本试剂盒检测了不同的衣原体,以及亲缘关系较近或者较常见的22种细菌,均未发现非特异性信号,沙眼衣原体、肺炎衣原体(Chlamydia pneumoniae)和兽类衣原体均不能产生特异性信号;对18个临床样本(包括痰、喉拭子和支气管冲洗液)进行检测,获得8个阳性,灵敏度高于ELISA法(6个阳性),与临床诊断结果一致;对源于鸽的9个鼻拭子样本进行检测,获得3个阳性。本试剂盒适用于鹦鹉热衣原体族的检测。

 

 

 

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