Method for detecting microorganisms

a microorganism and detection method technology, applied in the field of detection methods, can solve the problems of insufficient sample amount, inability to detect a plurality of pathogens, and difficulty in distinguishing between infectious uveitis and non-infectious uveitis, so as to avoid human error in the detection operation.

Inactive Publication Date: 2020-12-10
NAT UNIV CORP TOKYO MEDICAL & DENTAL UNIV +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0050]According to the present invention, a step of extracting a nucleic acid from a sample (for example, an aqueous humor) is unnecessary, so that the sample is hardly lost in the step of detecting a microorganism. Therefore, it is possible to simultaneously and quickly detect a plurality of pathogenic microbes even with a small amount of sample. Furthermore, since the microorganism detecting step in the present invention is simple, it is possible to avoid human error in the detecting operation.

Problems solved by technology

However, the distinction between infectious uveitis and non-infectious uveitis may be difficult only by clinical findings, and there are cases where the symptoms become serious due to delay of diagnosis or improper treatment.
The amount of sample may be insufficient to detect a plurality of pathogens.

Method used

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  • Method for detecting microorganisms
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Examples

Experimental program
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Effect test

example 1

[0075][Analysis of 53 Infectious Uveitis-Positive Samples]

[0076]20 μL of anterior chamber fluid or vitreous of samples obtained from 53 patients with suspected infectious uveitis were mixed with 180 μL of PCR buffer. The composition of the PCR buffer after mixing was 0.05% (w / v) nonionic surfactant, 1.5 mM MgCl2, 35 mM KCl and 200 μM dNTP (dATP, dGTP, dCTP and dTTP), respectively. 20 μL of the obtained mixed solution was dispensed into each tube of an 8-strip tube strip containing the solid composition for PCR reaction. The solid composition for PCR reaction in the strip tube comprises a DNA polymerase, a fluorescent dye-labeled oligonucleotide probe for fluorescent detection of a PCR amplification product different in each tube, and one or two different PCR primer pairs. In addition, it contains one or two types of different PCR primer pairs. The combinations of PCR primer pairs were as follows.[0077](i) Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene detection primer pair an...

example 2

[0089][Analysis of Samples Diagnosed as Non-Infectious Uveitis]

[0090]FIG. 3 shows the results of 51 samples obtained from patients diagnosed with non-infectious uveitis measured by the real-time PCR (qPCR) method and the method of the present invention. All samples were negative by the real-time PCR (qPCR) method. They were also negative by the method of the present invention. That is, it was shown that the measurement results obtained by both methods match with each other.

[0091]Sequence Table

[0092]20190607 Sequence Table

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Abstract

The present invention relates to a method for simultaneously detecting a plurality of pathogens from biologically-derived samples, and a kit for carrying out the method. Specifically, the present invention relates to a method for simultaneously detecting a plurality of pathogens that cause infectious uveitis, one of eye infections from samples such as anterior chamber fluid or vitreous by polymerase chain reaction (PCR), and a kit for carrying out the method.

Description

TECHNICAL FIELD[0001]The present invention relates to a method for simultaneously detecting a plurality of infectious microorganisms from biologically-derived samples, and a kit for carrying out the method. Specifically, the present invention relates to a method for simultaneously detecting a plurality of pathogens that cause infectious uveitis, one of eye infections from samples such as anterior chamber fluid or vitreous by polymerase chain reaction (PCR), and a reagent kit for carrying out the method.BACKGROUND TECHNOLOGY[0002]Uveitis is a general term for diseases that cause inflammation in the eye. In severe cases, it causes visual disorders such as blindness at a high rate. Uveitis is classified into non-infectious uveitis such as sarcoidosis, Harada disease and Behcet's disease, and infectious uveitis caused by a pathogen. The non-infectious uveitis is treated with an immunosuppressive drug such as a steroid drug. On the other hand, the infectious uveitis requires treatment wi...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/686C12Q1/70C12Q1/689C12Q1/6893C12Q1/6806C12Q1/32C12Q1/6816G01N21/64
CPCC12Q1/689C12Q1/6816C12Q1/686C12Q1/32C12Q1/6806C12Q1/705C12Q1/6893G01N21/6486C12Q1/702C12Q1/701
Inventor SHIMIZU, NORIOTAKASE, HIROSHIMOCHIZUKI, MANABUTOMARU, YASUHIRONAKANO, SATOKOSUGITA, SUNAOSHIKATA, MASAMITSU
Owner NAT UNIV CORP TOKYO MEDICAL & DENTAL UNIV
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