The invention provides a primer group and a probe group capable of simultaneously detecting various influenza viruses. Sequences of the primer group are shown as SEQ ID NO:1-8, and sequences of the probe group are shown as SEQ ID NO:9-12; and the influenza viruses include influenza A virus, influenza B virus, seasonal influenza virus H1 sub-type and H3 sub-type. On the basis, a kit, which is capable of simultaneously detecting the various influenza viruses, is provided. According to the prime group, the probe group and the kit, direct amplification and multiplex PCR are combined; direct RT-PCR can be conducted on suspected influenza cases or epidemiological investigation throat swab samples; and in comparison with a conventional method, a nucleic acid extraction step is avoided, so that it is conducive to implementation of automatic integrated detection and to improvement of efficiency. The prime group, the probe group and the kit have obvious advantages in aspect of being applied to rapid discrimination and detection of people infected by the influenza viruses, and the invention is simple and convenient to operate, short in detection time and relatively low in detection cost. The prime group, the probe group and the kit are applicable to rapid detection at a site of epidemic situation; and the prime group, the probe group and the kit have a broad and practical application value.

Primer group and probe group capable of simultaneously detecting various influenza viruses and kit capable of detecting various influenza viruses

The invention provides a method of preparing a DNA probe, a kit, a method of detecting content of bacillus coli DNA in a biological product, and a method of determining a production process of a biological product. The method of preparing the DNA probe comprises the steps of (1) ultrasonic crushing cell genomic DNA of bacillus coli, wherein in the ultrasonic crushing treatment, the concentration of the cell genomic DNA of bacillus coli is 20 mcg/ml; and (2) incubating the product of ultrasonic crushing and a labeling reagent so as to acquire the DNA probe, wherein the DNA probe carries the label. The DNA probe prepared by the method can detect the volume of residual bacillus coli DNA in biological products (comprising middle sample, semi-finished product and finished product) exclusively,flexibly, quickly and accurately.

Method of preparing DNA probe, kit and application thereof

The invention relates to a quad fluorescence PCR method for quickly identifying A, B, H1N1-2009 and quarter H3 sub-influenza viruses and belongs to the technical field of bioengineering. The method comprises the following steps: respectively mixing forward and reverse primers (40muM) of four primers at volume ratio of 1:1 under the condition of the designed single primer and probe having typical amplification curve for the influenza viruses and mixing four probes (20muM) at volume ratio of 1:1, thereby using as the primer and probe used in the invention; using a manual extracting kit or a full automatic nucleic acid extracting instrument for extracting nucleic acid (RNA) of an influenza sample; performing Real-time PCR detection; and judging the positive or negative of corresponding A, B, H1N1-2009 and quarter H3 sub-influenza viruses of an influenza sample through the PCR amplification curve according to a FAM channel Ct value, a HEX channel Ct value, a TEXAS RED channel Ct value and a Cy5 channel Ct value.

Quad fluorescence PCR method for quickly identifying A, B, H1N1-2009 and quarter H3 sub-influenza viruses

The invention belongs to the technical field of biology, and establishes an influenza A virus fast typing and analyzing process. Influenza A viruses can be quicky and accurately typed, analyzed and subjected to genetic evolution research. The influenza A virus fast typing and analyzing process comprises three technical points as follows: (1) reference sequences which are suitable for carrying out parting of the influenza A viruses are screened; (2) an influenza A virus parting and sequence analyzing method is established; and (3) parallelization and automation of sequence analysis are realized. By the influenza A virus fast typing and analyzing process, epidemiological investigation and genetic variation studies, which are carried out by various pestilence prevention and control institutions and research institutes, of influenza viruses can be met.

Influenza A virus fast typing and analyzing process

The invention discloses a fluorescent PCR (Polymerase Chain Reaction) method for rapidly identifying type A, B and seasonal H3 subtype influenza virus and belongs to the technical field of bioengineering. The method disclosed by the invention comprises the following steps: respectively mixing forward and reverse primers (40uM) of three primers according to a volume ratio of 1:1 under the condition that the designed single primer and probe have a typical amplification curve on influenza virus, mixing three probes (20uM)) according to a volume ratio of 1:1 so as to serve as service primers and probes in the invention; extracting nucleic acid (RNA) of an influenza sample by using a manual extraction kit or a full-automatic nucleic acid extraction instrument; performing Real-time PCR detection; and respectively judging that the influenza samples are various positive or negative A, B and seasonal H3 subtype influenza viruses by virtue of a FAM channel Ct value, an HEX channel Ct value and a TEXAS RED channel Ct value.

Fluorescent PCR (Polymerase Chain Reaction) method for rapidly identifying type A, B and seasonal H3 subtype influenza virus

The invention belongs to the field of biotechnologies, and particularly relates to a multi-PCR-ELISA detection kit for human seasonal influenza viruses and an application method thereof. The detection kit disclosed by the invention is composed of a RT-PCR reaction system, an ELISA detection system, four target-gene (an influenza-A-virus M gene, a H1 subtype virus HA gene, a H3 subtype virus HA gene, and a B type virus NS gene) positive plasmids (Pa-m, Ph1-ha, Ph3-ha and Pb-ns), a negative quality control specimen and four target-gene specific primer probes. Specific amplification is performed by using specific primers labeled by using four sets of biotins through RT-PCR, an amplified product after being denatured is hybridized with a specific probe labeled by using digoxin, a hybridized product is enveloped with a streptavidin-enveloped 96-hole micro-plate, and an anti-digoxin antibody labeled by using horse radish peroxidase is added for carrying out detection through an ELISA method, so that a rapid, sensitive and specific typing detection kit for seasonal influenza viruses such as H1 and H3 subtypes and hepatitis B viruses is established. The invention relates to the application of four sets of specific primer probes in the clinical differential diagnosis of influenza virus infection and the typing authentication of influenza virus isolates.

Typing detection kit for human seasonal influenza viruses and application method thereof

The utility model discloses a portable small liquid nitrogen tank. The device comprises an anti-skid base, a support frame and a handle, an outer tank body is arranged on the antiskid base; a protective fence is arranged on the outer side of the outer tank body; an inner tank body is arranged in the outer tank body; a liquid nitrogen adsorption block is arranged in the inner tank body; the upper end of the nitrogen inlet pipe is connected with the sealing cover; the supporting frame is arranged on the inner side wall of the nitrogen inlet pipe, the supporting frame is arranged below the heat preservation layer, the buckle is connected with a sample frame in a clamped mode, the sample frame is arranged above the liquid nitrogen adsorption block, the lifting handle penetrates through the protective guard and is connected with a connecting block through a connecting shaft, and the connecting block is arranged on the outer side of the outer tank body. The portable small liquid nitrogen tank is provided with the anti-skid base and the protective guard, the anti-skid base can reduce sliding of the liquid nitrogen tank in the transportation process, the protective guard can prevent foreign objects from colliding with the liquid nitrogen tank body in the violent vibration process in the transportation process to damage the liquid nitrogen tank body, and the lifting handle facilitates carrying and using of the small liquid nitrogen tank.

Portable small liquid nitrogen container

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Typing detection kit for human seasonal influenza viruses and application method thereof

Portable small liquid nitrogen container