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

Active Publication Date: 2018-06-19
JIANGSU BIOPERFECTUS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Existing fluorescent PCR HPV detection products have the following disadvantages: 1. Existing fluorescent PCR products cannot achieve 18 high-risk HPV typing in a single tube, resulting in low detection throughput and cumbersome operations that are difficult to meet the requirements for large-scale screening

Method used

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  • Nucleic acid composition for detecting HPVs (Human Papilloma Viruses) as well as application and kit of nucleic acid composition
  • Nucleic acid composition for detecting HPVs (Human Papilloma Viruses) as well as application and kit of nucleic acid composition
  • Nucleic acid composition for detecting HPVs (Human Papilloma Viruses) as well as application and kit of nucleic acid composition

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a nucleic acid composition for detecting HPVs (Human Papilloma Viruses) as well as application and a kit of the nucleic acid composition, and relates to the technical field ofin-vitro diagnosis of virus nucleic acid detection. The nucleic acid composition for detecting the HPV comprises primers which are shown in SEQ ID NO.1 to 6 and primers which are shown in SEQ ID NO.7to 9. The nucleic acid composition disclosed by the invention is capable of simultaneously detecting HPV 16 and HPV 18 and parting the specificity of the HPV 16 and the HPV 18, meanwhile, quality control can also be carried out on a detecting process through an internal control beta-globin gene detection result, false negatives can be reduced, and the nucleic acid composition has the characteristics of high sensitivity and strong specificity.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis of viral nucleic acid detection, in particular to a nucleic acid combination for detection of human papillomavirus and its application and kit. Background technique [0002] For a long time, cytology screening has been the only or main method of cervical cancer screening. Since the introduction of traditional Pap cytology testing into clinical practice in 1941, the incidence and mortality of cervical cancer have been greatly reduced, especially the mortality rate has been reduced by at least 70%. , but the accuracy of Pap cytology smears is affected by many factors, and the false negative rate is relatively high, ranging from 5% to 40%. In the past 30 years, cytological screening has experienced a revolution in detection technology and diagnostic methods, that is, the application of liquid-based cytology technology and computer-aided cell detection system. The former has significantly i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/708C12Q2545/101C12Q2563/107
Inventor 王河清张琳李秀林张蓉严浩荣涂小宝徐加发刘中华王国强
Owner JIANGSU BIOPERFECTUS TECH CO LTD
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