Real-time fluorescence RPA detection kit for aspergillus fumigatus as well as special primer and probe thereof

Abstract

The invention relates to a real-time fluorescence RPA detection kit for aspergillus fumigatus as well as a special primer and a probe of thereof, and particularly provides a primer composition for RPAdetection, the primer composition comprises a forward primer and a reverse primer, the forward primer has a nucleotide sequence shown as SEQ ID NO: 1, and the reverse primer has a nucleotide sequenceshown as SEQ ID NO: 2. The primer composition for RPA detection can be used for rapidly, conveniently, efficiently, specifically and sensitively detecting aspergillus fumigatus components in samplesto-be-detected, such as thalli, sputum, blood and bronchoalveolar lavage fluid, under an isothermal condition.

Description

technical field [0001] The invention relates to the field of biological detection, in particular, the invention relates to a real-time fluorescent RPA detection kit for Aspergillus fumigatus and special primers and probes thereof. Background technique [0002] Aspergillus fumigatus is a common opportunistic pathogen that is widely distributed in the environment. Its spores can easily cause invasive aspergillosis after being inhaled by immunocompromised or immunocompromised patients. Because its pathogenic mechanism is still unclear, There is no effective treatment, and the fatality rate is as high as 90%. The main fungus that causes invasive aspergillosis is Aspergillus fumigatus. Generally, the infection of Aspergillus fumigatus can only be detected in the late stage of the disease or after death. Therefore, early diagnosis and prevention of Aspergillus fumigatus is very important. In the past few decades, with the high-intensity immunosuppressive treatment and the widespr...

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Problems solved by technology

However, due to the patient's own low immunity and antifungal treatment, false negative or false positive results may be produced by serological testing [SuLahian A, Touratier S, Ribaud P. False positive test for aspergillus antigenemia related to concomitant administration of piperacillin and tazobactam. N Engl J Med.2003,349:2366-2367.]

The detection method of molecular biology mainly detects the specific genes of Aspergillus fumigatus through PCR technology (including fluorescent quantitative PCR). PCR technology has the advantages of high specificity, accuracy, sensitivity, and rapidity. The requirements are very high, the equipment is expensive, and strict temperature control is required, so it is difficult to apply to the clinical detection of Aspergillus fumigatus [Ostrosky-Zeichner L.Invasivemycoses: diagnostic challenges.Am J Med.2012,125:S14-24.]

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