Kit for detecting pathogenic bacteria of mucor lungs as well as use method and application of kit

A technology of mucormycosis and kits, applied in the direction of microbe-based methods, biochemical equipment and methods, microbiological determination/testing, etc., can solve the problem of lack of universality of primers

Active Publication Date: 2021-04-30
上海捷诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this primer combination has high specificity and high sensitivity, it can realize simple, rap...

Method used

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  • Kit for detecting pathogenic bacteria of mucor lungs as well as use method and application of kit
  • Kit for detecting pathogenic bacteria of mucor lungs as well as use method and application of kit
  • Kit for detecting pathogenic bacteria of mucor lungs as well as use method and application of kit

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 mucor lungs as well as a use method and an application of the kit. The kit of the present invention comprises a universal primer capable of being present in the form of a mixture, wherein the universal primer is capable of binding to a common gene conserved region of rhizopus oryzae, rhizomucor micranthum, aureobasidium pullulans, rhizopus stolonifer, and Cunninghamia lanceolata. The kit provided by the invention can realize one-time detection of various mucor fungi, the detection time does not exceed 2 hours, and the specific type of mucor can be further determined in one-time 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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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 上海捷诺生物科技有限公司
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