Nucleic acid composition, kit and detection method for detecting infectious pathogens of cat digestive tract

A nucleic acid composition and detection method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of single pathogen detection, easy pollution, long detection cycle, etc., and achieve high detection sensitivity and detection The effect of low limit and short detection time

Pending Publication Date: 2021-06-04
上海基灵生物科技有限公司
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used clinical diagnostic methods are mostly immunofluorescence antibody detection, serological detection, virus isolation and identification and other technologies, but there are still shortcomings such as low sensitivity, long detection cycle and single pathogen detection
Ordinary PCR method can specifically amplify the gene fragments of the virus with high sensitivity, but it needs to be analyzed by gel electrophoresis after amplification, which is easy to cause pollution

Method used

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  • Nucleic acid composition, kit and detection method for detecting infectious pathogens of cat digestive tract
  • Nucleic acid composition, kit and detection method for detecting infectious pathogens of cat digestive tract
  • Nucleic acid composition, kit and detection method for detecting infectious pathogens of cat digestive tract

Examples

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Embodiment 1

[0056] This embodiment provides a nucleic acid composition for detecting pathogens in the digestive tract of cats. It includes: a first reagent for detecting feline panleukopenia virus and a second reagent for detecting feline coronavirus, the first reagent includes primer pair 1 and probe 1, and the second reagent includes primer pair 2 and probe 2. The sequence of primer pair 1 is shown in SEQ ID NO.1-2, the sequence of probe 1 is shown in SEQ ID NO.3, the sequence of primer pair 2 is shown in SEQ ID NO.4-5, the probe The sequence of 2 is shown in SEQ ID NO.6. The specific sequence is shown in Table 1:

[0057] Table 1 Sequence information.

[0058]

Embodiment 2

[0060] This example provides a specific method for designing and screening the nucleic acid composition obtained in Example 1.

[0061] The inventor selected the strains whose sequence numbers are KX900570.1 (feline panleukopenia virus) and DQ848678.1 (feline coronavirus) respectively to carry out primer design by inquiring about feline panleukopenia virus and feline coronavirus. , entrusted Xi'an Qingke Biotechnology Co., Ltd. to synthesize the plasmid of the relevant pathogen, and use the primer probe design software to select the appropriate sequence, that is, try to avoid the dimer and hairpin structure of the primer, and the annealing temperature of the primer is about 60°C.

[0062] The designed primer sequences are shown in Table 2. In Table 2, two nucleic acid combinations are designed corresponding to each pathogen, and each nucleic acid combination includes a primer pair and a probe.

[0063] Table 2 The designed primers and probe sequences.

[0064]

[0065] ...

Embodiment 3

[0076] This example provides a kit for detecting feline digestive tract pathogens (FCoV, FPV), which includes the nucleic acid composition of Example 1.

[0077] The kit in this example is a 25 μL reaction system. In other embodiments, the system of the above kit can also be adaptively adjusted according to needs, for example, a 15 μL or 20 μL reaction system can be selected.

[0078] The 25 μL reaction system includes 12.5 μL of PCR reaction solution (including reverse transcriptase, Taq DNA polymerase, dNTP, MgCl, etc., PCR reaction solution from Bao Rui Biological Company), FCoV / FPV primer probe set (0.15 μL of FCoV-F , 0.2 μL of FCoV-R, 0.15 μL of FCoV-P, 0.2 μL of FPV-F, 0.2 μL of FPV-R, 0.15 μL of FPV-P), DEPC-treated water, and the template addition amount was set to 5 μL.

[0079] The PCR reaction conditions using the above kit are: (1) reverse transcription at 50°C for 5-30min; (2) pre-denaturation at 95°C for 2-5min; (3) denaturation at 95°C for 5-15s; (4) annealing ...

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Abstract

The invention discloses a nucleic acid composition, a kit and a detection method for detecting infectious pathogens of a cat digestive tract, and relates to the technical field of in-vitro diagnosis of molecular biotechnology. The invention provides the nucleic acid composition and the kit for detecting the infectious pathogens of the cat digestive tract. The nucleic acid composition and the kit thereof can realize rapid and accurate detection of cat leukopenia virus and cat coronavirus at most, which is of great significance for clinical diagnosis and treatment of cat digestive tract diseases. The nucleic acid composition and the kit provided by the invention have the technical advantages of high detection sensitivity, good specificity and low detection limit, are short in detection time, and are beneficial to early diagnosis and treatment of diseased cats.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis of molecular biotechnology, in particular to a nucleic acid composition, a kit and a detection method for detecting infectious pathogens in the digestive tract of cats. Background technique [0002] Feline gastrointestinal infectious disease is a common clinical disease in cats, characterized by varying degrees of gastrointestinal symptoms. Feline panleukopenia virus (Feline Panleukopenia Virus, FPV) and feline coronavirus (FelineCoronavirus, FCoV) are common pathogens that clinically harm the digestive tract of cats. Among them, FPV is often accompanied by: fever (above 40°C), diarrhea, dehydration and obvious leukopenia in cats, etc., high mortality, the disease is highly contagious and has various ways of detoxification, sick cats, recovered cats and invisible infection All infected cats will excrete toxins through excrement and secretions; FCoV infection is very common in cats, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2563/107C12Q2521/107
Inventor 王佳琳孙依欣张晓婷
Owner 上海基灵生物科技有限公司
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