RT(Reverse Transcription)-PCR (Polymerase Chain Reaction) kit for detecting parainfluenza virus in one step
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An RT-PCR and detection kit technology, applied in the field of one-step RT-PCR detection kits for parainfluenza virus, can solve the problems of human parainfluenza virus infection without vaccine, and achieve short detection time period, high specificity and detection efficiency high effect
Inactive Publication Date: 2011-08-03
WUHAN UNIV
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[0006] There is no effective vaccine to prevent human parainfluenza virus infection; however, researchers are developing vaccines for human parainfluenza virus types 1 and 3
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Embodiment 1
[0067] 1. Example 1---One-step RT-PCR detection kit for parainfection virus
[0068] 12 throat swab samples from patients with suspected parainfection virus infection were detected, and 200 μl of each sample solution was taken for virus nucleic acid extraction.
[0069] 1. The composition of the kit:
[0070]This kit includes one-step 5xRT-PCR Buffer, positive control, negative control, four pairs of specific primers and Enzyme Mix;
[0071] 2. Design specific primers for parainfluenza virus type 1
[0072] 1F: 5'-CCGGTAATTTCTCATACCTATG-3',
[0073] 1R: 5'-CCTTGGAGCGGAGTTGTTAAG-3';
[0074] Design of specific primers for parainfluenza virus type 2
[0075] 2F: 5'-AACAATCTGCTGCAGCATTT-3',
[0076] 2R: 5'-ATGTCAGACAATGGGCAAAT-3';
[0077] Design of specific primers for parainfluenza virus type 3
[0078] 3F: 5'-CTCGAGGTTGTCAGGATATAG-3',
[0079] 3R: 5'-CTTTGGGAGTTGAACACAGTT-3';
[0080] Design of specific primers for parainfluenza virus type 4
[0081] 4F: 5'-CTGAACGGT...
Embodiment 2
[0115] The above method was used to test the sputum samples of another 20 patients with suspected parainfluenza virus infection, and 8 samples were found to be positive for parainfluenza virus, including 2 cases of parainfluenza virus type 1 positive, 2 cases of parainfluenza virus type 2 positive, and parainfluenza virus positive in 2 cases. Three cases were positive for type 3 and one case was positive for parainfluenza virus type 4, the detection rate was 100%.
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Abstract
The invention discloses a RT (Reverse Transcription)-PCR (Polymerase Chain Reaction) kit for detecting parainfluenza virus in one step, and relates to a PCR kit in the technical field of biology. The kit comprises one-step 5xRT-PCRBuffer, a positive control substance, a negative control substance, four pairs of specific primers and EnzymeMix, wherein the sequences of the specific primers 1F and 1R designed for the parainfluenza virus A are respectively 5'-CCGGTAATTTCTCATACCTATG-3' and 5'-CCTTGGAGCGGAGTTGTTAAG-3'. The kit has the advantages of short detection time period, high detection efficiency, high specificity of the detected virus, high accuracy, simple operation, easy generalization and good repeatability of the experimental result; and the kit can carry out virus quantitative analysis, the detection sensitivity of the kit is higher than that of an immunology detection method.
Description
technical field [0001] The invention relates to a PCR kit in the field of biotechnology, in particular to a one-step RT-PCR (reverse transcription-polymerase chain reaction, reverse transcription-polymerase chain reaction) detection kit for parainfluenza virus. Background technique [0002] Parainfluenza virus belongs to the Paramyxoviridae family, a single-stranded RNA enveloped virus, and it and rhinoviruses, coronaviruses, adenoviruses and other viruses are considered to be the pathogens that cause the common cold. There are four serotypes of parainfluenza virus: type 1 can cause severe laryngitis (pseudomembranous laryngitis), tracheitis or bronchitis; Serious infection cases can only occur when children are 2 to 4 years old; type 3 is the pathogen of infantile bronchitis and pneumonia after respiratory syncytial virus, often causing infantile bronchiolitis and pneumonia, and can also cause pseudomembranous Laryngitis; type 4 causes milder symptoms and mostly causes upp...
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