Nucleic acid composition for detecting HPVs (Human Papilloma Viruses) as well as application and kit of nucleic acid composition
A kit and papilloma technology, applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of cumbersome operation, low detection throughput, and difficulty in meeting the requirements of large-scale screening, etc. Achieve the effects of avoiding non-specific amplification, high specificity and sensitivity
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Embodiment 1
[0052] The kit for detection of human papillomavirus provided in this embodiment includes the following components:
[0053] (1) PCR reaction solution:
[0054] The PCR reaction solution consists of RNase-free water, 10× PCR buffer, 25mM Mg 2+ , 10mM dNTPs composition (see Table 1).
[0055] Table 1 Composition of PCR reaction solution
[0056] Reagent name
Addition volume (μl) / 50 reaction
RNase-free water
357.5
10×PCR buffer
125
25mM Mg 2+
250
10mM dNTPs
37.5
total
770
[0057] (2) Enzyme mixture
[0058] The enzyme mixture includes DNA polymerase and reverse transcriptase (Table 2).
[0059] The composition of table 2 enzyme mixture
[0060] Reagent name
Addition volume (μl) / 50 reaction
Taq enzyme
29
UDG enzyme
1
total
30
[0061] (3) HPV reaction solution
[0062] Described HPV reaction liquid comprises following composition components:
[0063] Upst...
experiment example 1
[0095] When performing ultra-multiplex PCR reactions, a large number of primers and probes are added to a single reaction tube, which promotes the random combination of primers and probes in the polymerization reaction to generate a large number of non-specific amplification products, which seriously affects the sensitivity of the kit. Therefore, this experiment uses similar multi-tube typing HPV detection products on the market to evaluate the consistency, sensitivity and specificity of the kit in Example 1 in clinical application.
[0096] The human papillomavirus nucleic acid typing test kit (referred to as HPV21 typing kit) of Jiangsu Shuoshi Biological Co., Ltd. was selected as the reference kit for this evaluation. A total of 550 cervical exfoliated cell samples collected by the hospital were used. Consistency analysis was performed on the kit of Example 1.
[0097] 1HPV16 test results
[0098] The specific results detected by the kit of Example 1 are shown in Table 5. ...
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