LAMP (loop-mediated isothermal amplification) kit for detection of mycoplasma pneumoniae and special LAMP primer for detection of mycoplasma pneumoniae

A technology of Mycoplasma pneumoniae and a kit, which is applied in the biological field, can solve problems such as no Mycoplasma pneumoniae, and achieve the effects of high specificity, high specificity and high sensitivity

Inactive Publication Date: 2016-01-13
INST OF PLA FOR DISEASE CONTROL & PREVENTION +1
View PDF5 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no LAMP kits and special primers for the detection of Mycoplasma pneumoniae in China

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • LAMP (loop-mediated isothermal amplification) kit for detection of mycoplasma pneumoniae and special LAMP primer for detection of mycoplasma pneumoniae
  • LAMP (loop-mediated isothermal amplification) kit for detection of mycoplasma pneumoniae and special LAMP primer for detection of mycoplasma pneumoniae
  • LAMP (loop-mediated isothermal amplification) kit for detection of mycoplasma pneumoniae and special LAMP primer for detection of mycoplasma pneumoniae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1. Primer Design for LAMP Detection of Mycoplasma pneumoniae

[0041] The Mycoplasma pneumoniae P1 gene (GenBank number: CP002077.1) was retrieved from the NCBI nucleic acid database GenBank (http: / / www.ncbi.nlm.nih.gov), and the homology analysis was performed by BLAST software to obtain the Mycoplasma pneumoniae P1 gene The specific conserved target sequence (SEQIDNO: 1 in the sequence table), and then according to the conserved target sequence, use the software PrimerdesignV4 (http: / / primerexplorer.jp / e / ) to design primers for LAMP detection of Mycoplasma pneumoniae, design Five sets of primer combinations (code-named MP-4, MP-16, MP-22, MP-27, MP-50) were obtained. Through experimental comparison, the primer combination MP-16 was finally selected. The primer sequences are shown in Table 1.

[0042] Table 1 is used for carrying out the primer combination MP-16 that LAMP detects to Mycoplasma pneumoniae

[0043]

[0044]

Embodiment 2

[0045] Example 2, LAMP detection of Mycoplasma pneumoniae

[0046] 1. Determine the best primer combination for LAMP detection of Mycoplasma pneumoniae

[0047] Five sets of primer combinations (code-named MP-4, MP-16, MP-22, MP-27, MP-50) designed in Example 1 are used for Mycoplasma pneumoniae (derived from the Infectious Disease Control Institute of the Chinese People's Liberation Army). Center) for LAMP detection to obtain the best primer combination, the specific method is as follows:

[0048] 1) Genomic DNA extraction of Mycoplasma pneumoniae

[0049] Extract the genomic DNA of Mycoplasma pneumoniae with QIAampviralDNAminikit (Qiagen, Hilden, Germany) commercialized DNA extraction kit, and specific extraction method comprises the following steps:

[0050] 1. Pipette 560 μL of the prepared AVL buffer-carrierRNA (carrier RNA, provided by the QIAampviral DNA minikit kit) into a 1.5mL centrifuge tube (adjust the AVL buffer-carrierRNA according to the actual amount of the s...

Embodiment 3

[0082] Embodiment 3, the specific detection of the LAMP detection method of Mycoplasma pneumoniae of the present invention

[0083] Staphylococcus aureus (Streptococcusaureus), Klebsiella pneumoniae (Klebsiellapneumoniae), Neisseria meningitides (neisseriameningitides), H7N9 subtype avian influenza virus (ThebirdfluvirusofH7N9), Stenotrophomonas Maltophilia (Stenotrophomonas Maltophilia), Listeria monocytogenes, Haemophilus influenzae, Shigellasonnei, salmonella, corynebacterium diphtheriae, Mycobacterium Tuberculosis, Acinetobacter baumannii, Methicillin-resistant Staphylococcus (methicillin-resistantstaphylococcus), Escherichia coli (escherichia coli), negative control (double distilled water) and positive control (Mycoplasma pneumoniae (Mp) standard strain) (the above strains are from the Chinese People's Liberation Army Disease Control and Prevention The genomic DNA of the Infectious Disease Control Center) is the sample to be tested, and the LAMP detection method in the L...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a LAMP (loop-mediated isothermal amplification) kit for detection of mycoplasma pneumoniae and a special LAMP primer for detection of mycoplasma pneumoniae. The special LAMP primer for detection of mycoplasma pneumoniae is designed according to a specificity conservative target sequence of a mycoplasma pneumoniae P1 gene (GenBank number: CP002077.1). The LAMP primer is formed by six primers including outer primers MP-16F3 and MP-16B3, inner primers MP-16FIP and MP-16BIP and loop primers MP-16LF and MP-16LB. By the aid of the LAMP kit and the special LAMP primer for detection of mycoplasma pneumoniae, quickness, convenience, high efficiency, high specificity and high sensitivity in qualitative detection of the mycoplasma pneumoniae in samples of pure bacteria, sputum, bronchoalveolar lavage fluid, throat swabs and the like can be realized without complicated instruments, and a new technical platform is provided for detection of the mycoplasma pneumoniae.

Description

technical field [0001] The invention belongs to the molecular biological detection of bacteria in the field of biotechnology, in particular to a LAMP kit for mycoplasma pneumoniae and its special primer and its application in detecting mycoplasma pneumoniae. Background technique [0002] Mycoplasma pneumoniae (Mycoplasmapneumoniae, Mp) is one of the most common pathogens of respiratory tract infection. Epidemiological surveys in my country show that Mp has become the primary pathogenic bacteria of adult community-acquired pneumonia in my country. Mp infection can not only cause respiratory diseases, but also systemic multi-organ lesions, such as encephalitis, myocarditis, hepatitis, nephritis, immune hemolytic anemia, etc., and the number of critical cases caused by Mp has been increasing in recent years. Due to lack of cell wall, Mp is ineffective against commonly used antibiotics such as β-lactams. Therefore, timely identification of Mp infection is of great significance...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/35
CPCC12Q1/6844
Inventor 袁静柏长青苑鑫刘威牛文凯崔茜李璞媛胡璇李环冯羽中郑敬
Owner INST OF PLA FOR DISEASE CONTROL & PREVENTION
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap