Composition, kit and method for detecting pathogens causing respiratory tract infection and identifying pathogen species and application
A respiratory and composition technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of complex pathogens, poor detection rate and accuracy, achieve high throughput and avoid false positives and environmental pollution, reducing the effect of psychological burden
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Embodiment 1
[0064] Embodiment 1, primers and probes used in the present invention
[0065] The primers and probes used in the present invention are shown in Table 1 below (SEQ ID NO:1~24 only corresponds to the base sequence, and the labeling method here is only a specific example):
[0066] Table 1
[0067]
[0068]
Embodiment 2
[0069] Embodiment 2, detect the pathogen that causes respiratory tract infection and identify the method for the type of pathogen
[0070] The detection samples of the present invention are throat swabs, sputum, alveolar lavage fluid, and blood. The magnetic bead method is used to extract viral nucleic acid, and the following operations are performed in the sample processing room:
[0071] 2.1 Take an appropriate amount of 1.5mL sterilized centrifuge tubes, mark the negative control, positive control and samples to be tested respectively, and add 300 μL RNA extraction solution 1 to each tube;
[0072] 2.2 Add 200 μL of the sample to be tested or negative control or positive control to each tube; cover the tube cap, vortex and mix for 10 seconds, and centrifuge briefly;
[0073] 2.3 Add 100 μL RNA Extraction Solution 2-mix to each tube (mix well and draw), shake and mix for 10 seconds, then let stand at room temperature for 10 minutes;
[0074] 2.4 Place the centrifuge tube o...
Embodiment 3
[0094] Embodiment 3, the detection rate of composition of the present invention
[0095] Use the compositions in Table 1 of the present invention to detect according to the method described in Example 2, and design a series of controls in addition (other dual-probe schemes: probe 1: CY5-CTAACCTTAACCCCGCAGC-BHQ2; probe 2: CY5-AATCCTAACCATGGAGCAAG- BHQ2) is also detected. The test results are shown in Table 2 and Table 3.
[0096] Table 2. Statistical table of Ct value results of different rhinovirus detection schemes
[0097]
[0098] Note: The larger the Ct value, the worse the detection effect, and No Ct means negative.
[0099] It can be seen from the test results in the above table that enteroviruses (Coxsackievirus EV71, Coxsackievirus A6) will not be detected by using this patented solution for detection; at the same time, for rhinovirus positive samples, using this patented solution for detection, its Ct value Both are small, indicating that its detection effect is...
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