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Nucleic acid combination for detecting Fusobacterium nucleatum in night soil, and applications and kits thereof

A fusobacterium nucleatum and kit technology, applied in the field of biomedical testing, can solve the problems of unreported detection methods and inability to carry out quantitative detection, etc., achieve high sensitivity and specificity, convenient extraction, and simple operation

Pending Publication Date: 2017-04-26
WUHAN AIMISEN LIFE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the application of Fusobacterium nucleatum is mainly focused on oral diseases and common cancers; quantitative detection cannot be performed, and there are no reports on Fusobacterium nucleatum in feces and its specific detection methods

Method used

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  • Nucleic acid combination for detecting Fusobacterium nucleatum in night soil, and applications and kits thereof
  • Nucleic acid combination for detecting Fusobacterium nucleatum in night soil, and applications and kits thereof
  • Nucleic acid combination for detecting Fusobacterium nucleatum in night soil, and applications and kits thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] This embodiment provides a nucleic acid combination for detecting Fusobacterium nucleatum in feces. The nucleic acid combination includes a primer combination and a probe; the primer combination includes a first primer pair, a second primer pair and a third primer pair, and the base of the first primer pair The base sequence is as shown in SEQID NO.1-2, the base sequence of the second primer pair is as shown in SEQ ID NO.3-4, and the base sequence of the third primer pair is as shown in SEQ ID NO.5-6. The sequence of a primer pair is as follows:

[0079] Upstream primer SEQ ID NO.1:

[0080] 5'-TCTCCTTTCAATAAA AGTGGC-3';

[0081] Downstream primer SEQ ID NO.2:

[0082] 5'-AAGAAGTAGGATGAAAATCAGTTA-3';

[0083] The sequence of the second primer pair is as follows;

[0084] Upstream primer SEQ ID NO.3:

[0085] 5'-CCTGGACGAAGACTGACGCTC-3'

[0086] Downstream primer SEQ ID NO.4:

[0087] 5'-CTCAAGGGCACAACCTCCAAG-3';

[0088] The sequence of the third primer pair is ...

Embodiment 2

[0107] This embodiment provides a kit for detecting Fusobacterium nucleatum, which includes the nucleic acid combination provided in Example 1.

[0108] Of course, this kit also includes reagents that can directly perform PCR reactions, including PCR reaction buffer, TaqDNA polymerase, dNTPs and Mg 2+ one or more of.

[0109] For the method of using the kit, refer to the PCR amplification reaction system and the PCR reaction procedure provided in Example 1, the difference is that during the PCR reaction, the combined temperature for annealing and extension is 65°C.

Embodiment 3

[0111] This embodiment provides a kit for detecting Fusobacterium nucleatum, which includes the combination of detection primers for detecting Fusobacterium nucleatum as provided in Example 1.

[0112] The kit provided in this embodiment also includes a sample preservation solution, which includes 0.1 mol / L EDTA, 3 mol / L Tris·HCl and 0.15 mol / L NaCl.

[0113] Of course, this kit also includes reagents that can directly perform PCR reactions, including PCR reaction buffer, TaqDNA polymerase, dNTPs and Mg 2+ one or more of.

[0114] For the method of using the kit, refer to the following steps:

[0115] 1.1 Stool collection Collect the patient’s stool into the sample preservation solution, store it at room temperature or 4°C for 2 weeks, and store it at -20°C for 6 months; add 100 μL proteinase K, 0.5 mL of nucleic acid lysate, mix well;

[0116] 1.2 Incubate at 70°C for 10 minutes;

[0117] 1.3 Add 100mg of PVPP for crude extraction to remove impurities such as pigment or p...

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Abstract

The invention discloses a nucleic acid combination for detecting Fusobacterium nucleatum in night soil, and applications and kits thereof, and belongs to the technical field of biomedical examination. The nucleic acid combination for detecting Fusobacterium nucleatum in night soil can specifically detect the Fusobacterium nucleatum. The nucleic acid combination is used to prepare a kit for detecting Fusobacterium nucleatum in night soil and a kit for detecting cancer. The kits effectively improve the detection precision, can be conveniently and fast applied to detect the Fusobacterium nucleatum, and have high sensitivity and specificity. A technologic method provided by the invention can be used to conveniently extract Fusobacterium nucleatum genome from the night soil, and has the advantages of quantitative detection of the Fusobacterium nucleatum, simplicity in operation, and facilitation of promotion and application.

Description

technical field [0001] The invention relates to the technical field of biomedical testing, and in particular to a nucleic acid combination for detecting Fusobacterium nucleatum in feces, its application and a kit. Background technique [0002] Colorectal cancer (CRC) is a common multiple malignant tumor of the digestive tract, and its morbidity and mortality rank third among common malignant tumors in the world. In China, the incidence of colorectal cancer has been increasing year by year in recent years. It is estimated that there are about 400,000 new cases each year, and it currently ranks second among malignant tumors of the digestive system in my country. The 5-year survival rate of patients with early colorectal cancer can be as high as 90%, while the 5-year survival rate of patients with intermediate and advanced stages is only about 10%. About 80% of clinically diagnosed colon cancers are advanced, which leads to no obvious mortality rate. One of the key factors for...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6886C12Q1/689C12Q2600/166
Inventor 张良禄董兰兰姚希辉
Owner WUHAN AIMISEN LIFE TECH CO LTD
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