Unlock instant, AI-driven research and patent intelligence for your innovation.
Specific primer and kit for detecting FAV-8 and FAV-11
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A FAV-11, FAV-8 technology, applied in DNA/RNA fragments, microorganisms, recombinant DNA technology, etc., can solve the problem of indistinguishable virus infection, and achieve the effect of good specificity and high sensitivity
Active Publication Date: 2019-12-27
POULTRY INST SHANDONG ACADEMY OF AGRI SCI
View PDF4 Cites 0 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0003] Aiming at the above-mentioned prior art, in order to solve the clinically indistinguishable problem of chicken inclusion body hepatitis, the two kinds of viral infections, the present invention provides a method for detecting and distinguishing avian adenovirusserotype 8 (FAV-8) and avian adenovirus. Specific primers and kits for virus serotype 11 (FAV-11)
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0042] The design of embodiment 1 detection primer
[0043] According to the 52K gene sequences of group I avian adenovirus types 8 and 11 registered in Genbank, multiple sets of primers were designed using Oligo 6.0 software, and the primer FAV8-RTF4 with the best sensitivity, specificity and effect was screened out through experiments / FAV8-RTR4 and FAV11-RTF1 / FAV11-RTR1 are used for the identification and detection of FAV-8 and FAV-11, respectively.
[0044] Design primers for FAV-8:
[0045] FAV8-RTF1: 5'-GAAGGACAGCAGCAGCGTCGTCT-3';
[0046] FAV8-RTR1: 5'-CGGCTTGATGATGTCGATA-3';
[0047] The size of the amplified fragment is 87bp;
[0048] FAV8-RTF2: 5'-AAGGACAGCAGCGTCGTCT-3';
[0049] FAV8-RTR2: 5'-ACTCCGGCTCGGTGGCT-3';
[0050] The size of the amplified fragment is 134bp;
[0051] FAV8-RTF3: 5'-CACCAGCAGCGCTATCGACA-3';
[0052] FAV8-RTR3: 5'-GGGCGGACGCGACTC-3';
[0053] The size of the amplified fragment is 86bp;
[0054] FAV8-RTF4: 5'-AAGGACAGCAGCGTCGTCT-3';
...
Embodiment 2
[0074] The optimization of embodiment 2 dual fluorescent quantitative PCR reaction conditions
[0075] The reaction conditions were optimized by adjusting the primer concentration, annealing temperature, and reaction time. The optimized reaction conditions were: TB Green Premix Ex TaqⅡ10μL; 10μM FAV8-RTF4 0.3μL; 10μM FAV8-RTR40.3μL; 10μM FAV11-RTF1 1μL; 10μM FAV11-RTR1 1μL; 2 O 5.4 μL for a total volume of 20 μL. Placed in Roche LightCycler 96 fluorescent quantitative PCR amplification instrument for reaction, reaction program: 95°C 60sec pre-denaturation; 95°C denaturation 10sec, 58°C annealing 10sec, 72°C extension 15sec, a total of 40 cycles; melting curve 95°C 10sec, 60 seconds at 65°C, 1 second at 97°C.
Embodiment 3
[0076] The specificity of embodiment 3 double fluorescence quantitative PCR
[0077] Extract avian adenovirus serotypes 1, 4, 8, and 11, avian influenza virus (AIV), Newcastle diseasevirus (NDV), and avian reticuloendotheliosis virus (REV) according to the instructions of the viral nucleic acid extraction kit. , chicken anemia virus (CAV), chicken Marek's disease virus (MDV), chicken infectious bursal virus (IBDV), chicken infectious bronchitis virus (IBV), chicken infectious laryngotracheitis virus (ILTV) Nucleic acid, and use this as a template to carry out double fluorescence quantitative PCR under the above reaction conditions, and judge the result by PCR amplification curve and melting curve Tm value. As a result, the dual fluorescent quantitative PCR method did not show specific amplification and melting curve peaks in the detection of ALV, NDV, REV, CAV, MDV, IBDV, IBV, ILTV, FAV-1 and FAV-4, and the results were all negative. However, there are two specific melting c...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More
PUM
Login to View More
Abstract
The invention discloses a specific primer for detecting FAV-8 and FAV-11, a kit for detecting the FAV-8 and the FAV-11, and a double fluorescence quantitative PCR method for detecting and identifyingthe FAV-8 and the FAV-11. The specific primer comprises a detection primer, for detecting the FAV-8, shown in SEQ ID NO.1 and 2, and a detection primer, for detecting the FAV-11, shown in SEQ ID NO.3and 4; the kit comprises the specific primer; and the double fluorescence quantitative PCR method comprises the steps: (1) pathologic specimen nucleic acid is extracted; (2) a PCR reaction is conducted; and (3) after amplification, an amplified product is detected, the FAV-8 and the FAV-11 are distinguished according to a double fluorescence quantitative PCR amplification curve and solution curveTm value, if the Tm value is 88.21-88.81 DEG C, the detection result is that the FAV-8 is positive, and if the Tm value is 93.15-93.80 DEG C, the detection result is that the FAV-11 is positive. The lowest detection amount of the FAV-8 and the FAV-11 is 31 copies / [mu]L and 34 copies / [mu]L, the advantages of high sensitivity, good specificity and the like are achieved, and important clinical application value is achieved.
Description
technical field [0001] The invention relates to a detection method for detection and identification of chicken inclusion body hepatitis infection of common serotypes, in particular to a method for detection and identification of avian adenovirusserotype 8 (FAV-8) and avian adenovirusserotype 11 (FAV-11). 11) The specific primer and kit belong to the technical field of biological detection. Background technique [0002] Fowl adenovirus (Fowl adenovirus, FAV) belongs to the genusAvian Adenovirus of the family Avidae Adenoviridae, and is divided into three groups, among which group I adenovirus contains 12 serotypes. Chicken inclusion body hepatitis (Inclusion body hepatitis in chicken, IBH) is an acute infectious disease of chickens caused by group I adenovirus infection. The morbidity rate can reach 10-20%, and the mortality rate is relatively low, generally no more than 5%, and returns to normal 5-7 days after the onset. The main manifestations of sick chickens are depr...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to View More 
Login to View More