A real-time fluorescent quantitative PCR kit for auxiliary diagnosis of tuberculous pleurisy and its application

Abstract

The invention discloses a real-time fluorescent quantitative PCR kit for assisting in the diagnosis of tuberculous pleurisy and its application. The kit contains primers and probes for detecting Mycobacterium tuberculosis specific small RNA fragments (comprising Mtb-ms-1 and Mtb-ms-2), wherein the sequence of Mtb-ms-1 is as SEQ ID NO.1 As shown, the sequence of Mtb-ms-2 is shown in SEQ ID NO.2. The present invention extracts the total RNA of a small amount of pleural effusion of patients with pleurisy, reverse-transcribes the specific small RNA fragment of Mycobacterium tuberculosis into complementary cDNA, and uses specific primer probes for real-time fluorescent quantitative polymerase chain reaction detection, and its diagnostic sensitivity and specificity The results were high, able to significantly distinguish tuberculous pleurisy from the control group, superior to other diagnostic methods such as Mycobacterium tuberculosis culture, acid-fast smear staining, and DNA testing.

Description

technical field [0001] The invention relates to a kit for detecting specific small RNA fragments encoded by Mycobacterium tuberculosis in pleural effusion of patients with tuberculous pleurisy based on real-time fluorescent quantitative polymerase chain reaction method, which is applied to the auxiliary diagnosis of tuberculous pleurisy. The invention belongs to the technical field of medicine. Background technique [0002] Tuberculosis is one of the infectious diseases with the highest morbidity and impact worldwide, and about 1 / 4 of it is extrapulmonary tuberculosis. Tuberculous pleurisy is the second most common cause of exudative pleurisy, second only to lymphatic tuberculosis. Traditional diagnostic methods for tuberculous pleurisy include Mycobacterium tuberculosis culture, acid-fast smear staining, pleural biopsy, etc. The diagnostic sensitivity is low and time-consuming, which often leads to delays in treatment timing. In recent years, the molecular detection techn...

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Problems solved by technology

[0004] The present invention overcomes the defects of low diagnostic sensitivity, long time consumption and high cost of existing laboratory diagnostic methods for tuberculous pleurisy, and provides a sensitive, specific and rapid molecular biological detection method, which has a high sensitivity for diagnosing tuberculous pleurisy. It is superior to traditional tuberculosis detection and tuberculosis DNA detection methods, so it can be used as a new diagnostic method for tuberculous pleurisy

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