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首页> 外国专利> Procedure for the clinical diagnosis of an infectious disease based on the use of quantitative PCR, labeled oligonucleotides of 8 to 9 nucleotides in length and the UNG of the Atlantic Cod (Gadus morhua).

Procedure for the clinical diagnosis of an infectious disease based on the use of quantitative PCR, labeled oligonucleotides of 8 to 9 nucleotides in length and the UNG of the Atlantic Cod (Gadus morhua).

机译：基于定量PCR，长度为8至9个核苷酸的标记寡核苷酸和大西洋鳕（UNGS）的UNG的临床诊断程序。

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摘要

The present invention provides a method, useful for use in a clinical microbiology laboratory, which allows the determination of the presence or absence of a microorganism in a Biological sample of a subject with a high degree of reliability using the chain reaction of the Polymerase (PCR) performed quantitatively in real time. Said procedure comprises the following steps: #a. Carry out a treatment of the reaction mixture that will be used to carry out the polymerase chain reaction (PCR) conducted quantitatively in real time with the Uracil-DNA-Glycosylase from the Atlantic Cod (Gadus morhua) of SEQ ID No 3 or with a variant thereof; and # b. Determine the presence or absence of the microorganism in the biological sample, after treatment of the reaction mixture according to section a), by determining the presence or absence of amplicons from the polymerase chain reaction (PCR) performed quantitatively in real time; where said polymerase chain reaction (PCR) uses DNA, complementary DNA (cDNA) or single stranded DNA obtained by re-transcription of ribonucleic acid (RNA) from the biological sample, at least one pair of specific primers, dNTPs, a suitable reaction buffer, a thermostable DNA polymerase and a fluorescently labeled probe, wherein said fluorescently labeled probe consists of an oligonucleotide with a length of 8 to 9 nucleotides labeled with a fluorophore, preferably said probe has a length of 8 nucleotides.
机译：本发明提供了一种可用于临床微生物学实验室的方法，其允许使用聚合酶（PCR）的链反应以高度的可靠性确定受试者的生物学样品中微生物的存在或不存在。 ）实时定量执行。所述过程包括以下步骤：#a。用来自SEQ ID No 3的Atlantic Cod（Gadus morhua）的Uracil-DNA-糖基化酶或以SEQ ID No.3进行实时定量反应的聚合酶链反应（PCR）的反应混合物的处理。其变体;和＃b。在根据a）部分处理反应混合物后，通过实时定量检测聚合酶链反应（PCR）中扩增子的存在与否，确定生物样品中是否存在微生物;其中所述聚合酶链反应（PCR）使用DNA，互补DNA（cDNA）或通过从生物样品中核糖核酸（RNA）重新转录而获得的单链DNA，至少一对特异性引物，dNTP，合适的反应缓冲液，热稳定的DNA聚合酶和荧光标记的探针，其中所述荧光标记的探针由长度为8至9个核苷酸的寡核苷酸组成，所述寡核苷酸被荧光团标记，优选地，所述探针的长度为8个核苷酸。

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