Reagent kit for detecting pathogenic bacteria of pulmonary mucormycosis and its use method and application

A technology for mucormycosis and pathogenic bacteria, which is applied in the field of fungal detection and can solve the problem of primers not being universal

Active Publication Date: 2021-10-15
上海捷诺生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this primer combination has high specificity and high sensitivity, it can realize simple, rapid and accurate detection, but this method uses multiple pairs of primers, and the primers are not universal

Method used

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  • Reagent kit for detecting pathogenic bacteria of pulmonary mucormycosis and its use method and application
  • Reagent kit for detecting pathogenic bacteria of pulmonary mucormycosis and its use method and application
  • Reagent kit for detecting pathogenic bacteria of pulmonary mucormycosis and its use method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] This embodiment is an exemplary mucormycetes detection kit, which is based on fluorescent quantitative PCR technology, and is designed to detect Rhizopus oryzae, Rhizomucor minimus, Rhizopus umbellatus, Rhizopus stoloniferus and Cinghamella thorns Wait.

[0036] The kit of this embodiment includes a PCR mixture (optimized), primers and fluorescent probes.

[0037] 3. The composition of the kit is shown in Table 1:

[0038] Table 1 Kit Composition

[0039] component name Specification (μl) Cap color Mucor PCR mix >250 amber hot start polymerase >25 Purple Diluent >950 transparent internal quality control >500 black positive control >125 White

[0040] Wherein, the PCR mixture contains 10 mM Tris-HCl (pH 8.8 at 25° C.), 50 mM KCl, 0.08% ethylphenyl polyethylene glycol, and the primers are primer pairs 1 and 2 shown in Table 2.

[0041] Quality controls included M13 as an internal control (IC) with a Ct value bet...

Embodiment 2

[0043] This embodiment is another exemplary mucormycetes detection kit, which is based on fluorescent quantitative PCR technology, and is designed to detect Rhizopus oryzae, Rhizomucor minimus, Rhizopus umbellatus, Rhizopus stoloniferus, and Rhizopus spinosa. mold, and identify its specific type. Except for the following description, the rest is the same as Example 1.

[0044] The kit of this embodiment includes an optimized composition of reaction solution, primers and fluorescent probes.

[0045] Among them, the PCR mixture contains 10mM Tris-HCl (pH 8.8at 25 ℃), 50mM KCl, 0.08% ethyl phenyl polyethylene glycol, 2mM MgSO 4 .

[0046] Wherein, the primer is the primer pair 1 shown in Table 1, and further includes a probe set, which is composed of five probes labeled with different fluorescence respectively, and nucleic acid analogs are introduced into probes 2, 4 and 5 to increase their Tm values .

[0047] Table 2 Primer and probe sequence and concentration

[0048] ...

Embodiment 3

[0052] This example is an experimental example using a kit.

[0053] 1. Storage conditions of samples and kits

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Abstract

The invention discloses a kit for detecting pathogenic bacteria of pulmonary mucormycosis and its use method and application. The kit of the present invention comprises a universal primer that can exist in the form of a mixture, wherein the universal primer can bind to the common gene of Rhizopus oryzae, Rhizomucor minimus, Rhizopus umbelliferum, Rhizopus stoloniferus and Cinghamella thorns Conserved area. The kit of the invention can realize one-time detection of multiple mucormycetes, and the detection time is not more than 2 hours, and can further determine the specific type of mucormycetes in one detection.

Description

technical field [0001] The invention relates to the field of fungal detection, in particular to a kit for detecting pathogenic bacteria of pulmonary mucormycosis and its use method and application. Background technique [0002] Mucormycosis is a rare and highly fatal (over 50%) invasive fungal infection characterized by vascular invasion and subsequent tissue necrosis by thrombosis. Mucormycosis mostly occurs in people with obvious risk factors or predisposing factors, including patients with organ transplantation, hematological malignancy, diabetes or renal failure, and is often covered up by underlying diseases, so the rate of clinical misdiagnosis and missed diagnosis is high. Mucormycosis progresses rapidly, and controlling the primary disease, antifungal treatment, and surgical removal of local lesions are the key to reducing the mortality rate. [0003] Due to the lack of laboratory test support, it is difficult to make a diagnosis based on clinical experience alone. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/785
CPCC12Q1/686C12Q1/6895C12Q2561/113
Inventor 黄迎燕杨晓明杜金凤夏小凯程鲁向朱丽媛吉斯·丁格曼斯
Owner 上海捷诺生物科技股份有限公司
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