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Nucleic acid composition and test kit for blood screening

A technology for nucleic acid composition and blood screening, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., to achieve the effect of reducing the risk of missed detection

Active Publication Date: 2020-05-29
ZHUHAI LIVZON DIAGNOSTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to public reports, there is no multiple nucleic acid detection kit capable of simultaneously detecting HBV / HCV / HIV-1 / HIV-2 / SARS-Cov-2

Method used

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  • Nucleic acid composition and test kit for blood screening
  • Nucleic acid composition and test kit for blood screening
  • Nucleic acid composition and test kit for blood screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The blood screening nucleic acid composition provided in this embodiment includes:

[0041] The first primer set is shown in SEQ ID NO.1 and SEQ ID NO.2, and the nucleic acid sequence of the first probe is shown in SEQ ID NO.3; wherein, the 5' end of the first probe is labeled with a fluorescent reporter Group FAM, the 3' end is labeled with a fluorescent quencher group BHQ1;

[0042] The second primer set is shown in SEQ ID NO.4 and SEQ ID NO.5, and the nucleic acid sequence of the second probe is shown in SEQ ID NO.6; wherein, the 5' end of the second probe is labeled with a fluorescent reporter Group FAM, the 3' end is labeled with a fluorescent quencher group BHQ1;

[0043] The third primer set is shown in SEQ ID NO.7 and SEQ ID NO.8, and the nucleic acid sequence of the third probe is shown in SEQ ID NO.9; wherein, the 5' end of the third probe is labeled with a fluorescent reporter Group CY5, the 3' end is labeled with a fluorescent quencher group BHQ1;

[0044...

experiment example 1

[0075] Verify the sensitivity of the nucleic acid composition of Example 1

[0076] The detection method is as follows:

[0077] HCV national reference product (340016-201701), HBV national reference product (300022-201601), HIV-1 WHO standard product (16 / 194), HIV-2 WHO standard product (16 / 296) and SARS-Cov-2 containing The positive plasmid of the ORF1ab gene fragment (SEQID NO.23) (its backbone is the pUC57 vector, see the vector map Figure 9 ) was diluted to the concentration in Table 5 for nucleic acid extraction (for the extraction method, refer to the extraction method of the nucleic acid sample to be tested in Example 1. In this experimental example, the ORF1ab plasmid of SARS-Cov-2 was extracted, mainly to simulate SARS-Cov-2 After the sample is extracted by the extraction method used in the present invention, the sensitivity of detection with the kit of the present invention), each concentration is 3 tubes, and 10 μL of internal control RNA pseudovirus (containing ...

experiment example 2

[0087] Verify the specificity of the nucleic acid composition of Example 1

[0088] Detection of human cytomegalovirus, hepatitis A virus, syphilis, herpes simplex virus type 1, herpes simplex virus type 2, Candida albicans, respiratory syncytial virus type A, respiratory virus type B, influenza A virus H1N1, influenza A Virus H3N2, influenza A virus H1N1 (2009), influenza B virus Yamagata and influenza B virus Victoria virus, and cultures of Staphylococcus aureus to verify whether there is cross-reaction with the nucleic acid composition of the example.

[0089] The specimens or cultures of the above 8 kinds of viruses and reference substances (HBV / HCV / HIV-1 positive control, HIV-2 positive control, SARS-Cov-2 positive control contain plasmids containing ORF1ab) according to Example 1 The extraction method was used for extraction, and 10 μL of internal control RNA pseudovirus was added to each tube, the extraction volume was 0.5 mL, and the elution volume was 60 μL. After ext...

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Abstract

The invention relates to the field of virus molecular inspection, and particularly provides a nucleic acid composition and a test kit for blood screening. The nucleic acid composition used in the blood screening provided by the invention comprises a nucleic acid composition for detecting HBV, HCV, HIV and COVID-2019. Researches of the inventor prove that the nucleic acid composition provided by the invention is strong in specificity, high in sensitivity and strong in interference resistance, and can realize rapid detection of HBV / HCV / HIV-1 / HIV-2 / SARS-Cov-2 in blood screening. Specific double-segment nucleic acid combination design is carried out on HIV-1 and HBV genomes, the situation of missing detection caused by virus gene mutation is greatly reduced, the detection rate is greatly increased, the residual risk of transfusion infection is reduced, screening is carried out on SARS-Cov-2 genomes, and the interference on the detection of HBV and HCV in the combination is obviously reduced. The blood screening detection test kit containing the nucleic acid combination provided by the invention has the advantages of high detection rate, good specificity and strong interference resistance.

Description

technical field [0001] The invention relates to the field of virus molecular detection, in particular to a nucleic acid composition for blood screening and a kit comprising the nucleic acid composition. Background technique [0002] With the improvement of people's living standards and the deepening of medical reform in our country, people's demand for medical treatment continues to increase, followed by a rapid increase in the demand for clinical blood. Blood can save a patient's life, but at the same time, transfusion of blood or blood products has the risk of spreading disease. The World Health Organization (WHO) has officially listed blood safety as one of the global priority health tasks. [0003] At present, blood safety testing mainly adopts immunology and clinical biochemical testing methods. The enzyme-linked immunosorbent reagent detects the antibodies produced by the human body after being infected by the virus, and the window period is long (from the time when t...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/701C12Q1/703C12Q1/706C12Q1/707C12Q2531/113C12Q2563/107Y02A50/30
Inventor 勾宏娜吴燕欧格宋冰燕邓京林艳
Owner ZHUHAI LIVZON DIAGNOSTICS
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