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Kit and detection method for synchronously detecting 19 encephalitis and meningitis pathogens based on RT-PCR and capillary electrophoresis

A technology of capillary electrophoresis and simultaneous detection, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection. Strong specificity and good repeatability

Pending Publication Date: 2020-07-28
NINGBO HEALTH GENE TECHNOLOGIES CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The complex composition of infectious disease specimens and the low concentration of pathogenic microorganism nucleic acid all restrict the NGS detection of pathogenic microorganisms in cerebrospinal fluid

Method used

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  • Kit and detection method for synchronously detecting 19 encephalitis and meningitis pathogens based on RT-PCR and capillary electrophoresis
  • Kit and detection method for synchronously detecting 19 encephalitis and meningitis pathogens based on RT-PCR and capillary electrophoresis
  • Kit and detection method for synchronously detecting 19 encephalitis and meningitis pathogens based on RT-PCR and capillary electrophoresis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] (1) Selected samples: cerebrospinal fluid samples from patients.

[0062] (2) Sample collection

[0063] Collection method: CSF samples were collected by lumbar puncture. The patient is placed in a supine position with the neck flexed and the hips and knees flexed to widen the intervertebral space. The puncture point is in the space between the 3rd and 4th lumbar vertebrae.

[0064] Valid sample: The appearance should be colorless and transparent liquid, the sample volume should be more than 1mL, and there should be no blood pollution.

[0065] (3) Nucleic acid extraction: 300 μL of cerebrospinal fluid sample was taken for nucleic acid extraction, and 80 μL of ME positive control and negative control were taken for extraction, and each sample involved in the extraction was extracted with 2 μL of IC.

[0066] (4) Preparation of reaction system

[0067] According to the instructions, prepare the reaction system according to the ratio of 4.6 μL of ME master mix and 0.4...

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Abstract

The invention discloses a kit and detection method for synchronously detecting 19 encephalitis and meningitis pathogens based on RT-PCR and capillary electrophoresis. The kit comprises an ME premixedsolution, an RT-PCR enzyme solution, an ME positive control, a negative control, a reaction internal reference (IC) and nuclease-free water. Specific gene nucleic acid sequences of the 19 encephalitisand meningitis pathogens and 3 internal references (a human DNA internal reference, a human RNA internal reference and the reaction internal reference) are amplified at the same time by adopting a one-step RT-PCR method, amplification products with different lengths are separated by adopting capillary electrophoresis, and detection results of the pathogens are obtained. The kit can synchronouslydetect the 19 encephalitis and meningitis pathogens and provide cross infection information, and has the advantages of high sensitivity, strong specificity, good repeatability, rapidness and accuracy.

Description

technical field [0001] The invention relates to the technical field of pathogen nucleic acid detection, in particular to a kit and a detection method for synchronously detecting 19 kinds of encephalitis and meningitis pathogens based on RT-PCR and capillary electrophoresis. Background technique [0002] Meningitis is an acute inflammation of the protective membranes that surround the brain and spinal cord, collectively known as the meninges. Inflammation of the meninges can be caused by infection with viruses, bacteria, other microorganisms, and in rare cases, certain drugs. Meningitis can be fatal because of the proximity of the inflammation to the brain and spinal cord; therefore, it is a medical emergency. [0003] The most common symptoms of meningitis are headache and neck stiffness, accompanied by fever, confusion or altered consciousness, vomiting, and sensitivity to light (photophobia) or loud sounds (phonophobia). Children usually only present with nonspecific sym...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/686C12R1/93
CPCC12Q1/701C12Q1/705C12Q1/689C12Q1/686C12Q2600/166C12Q2600/16
Inventor 吴勇曾县平余道福何秀才必华孔咪咪余丁
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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