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LAMP (loop-mediated isothermal amplification) kit for early diagnosis of candida krusei and special primer

A technology for early diagnosis of Candida krusei, applied in recombinant DNA technology, microbiological measurement/testing, biochemical equipment and methods, etc., can solve the problems of long time period for diagnosis of Candida krusei infection, and achieve convenient result interpretation , highly operable effect

Active Publication Date: 2016-09-21
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In summary, based on DNA sequence analysis, try to find a fast, simple, operable, and easy-to-interpret method for the rapid and early diagnosis of Candida krusei infection, which will provide a basis for clinical early diagnosis and timely treatment. Convenience, which is of great significance to solve the problems of long time period for the diagnosis of Candida krusei infection in clinical practice, high requirements for testing personnel skills and laboratory platform conditions

Method used

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  • LAMP (loop-mediated isothermal amplification) kit for early diagnosis of candida krusei and special primer
  • LAMP (loop-mediated isothermal amplification) kit for early diagnosis of candida krusei and special primer
  • LAMP (loop-mediated isothermal amplification) kit for early diagnosis of candida krusei and special primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Screening of primers for early diagnosis of Candida krusei infection and verification of its specificity and sensitivity

[0053] 1. Main experimental materials and equipment

[0054] 1. Strains

[0055] The standard and clinical strains of 6 strains of Candida krusei were selected, namely: ATCC(F)C6d, ATCC 2159, SCZ50016, SCZ50018, SCZ50019, SCZ50020, and other common clinical strains used to verify the specificity of the LAMP method. 20 standard strains and 20 clinical strains, all the strains come from the strain bank of Shanghai Key Laboratory of Molecular Biology of Medical Fungi, a total of 46 strains.

[0056] 2. Equipment

[0057] Ultra-clean bench: CA-1390-1 vertical laminar flow clean bench, Shanghai Shangjing Purification Equipment Co., Ltd.; biological safety cabinet: NUAIRE Biological Safefy Cabinet Class II; water-proof electric heating constant temperature incubator: Heraeus B5060EKCO 2 ; Pure water machine: HITECH water purification system; ...

Embodiment 2

[0154] Embodiment 2 is used for the test kit of early diagnosis candida krusei

[0155] The kit contains the following kit materials:

[0156] 1. Primer solution: ADE2-F3 (SEQ ID NO.6, 5 μmol / L), ADE2-B3 (SEQ ID NO.7, 5 μmol / L), ADE2-BIP (SEQ ID NO.9, 40 μmol / L), ADE2-FIP (SEQ ID NO.8, 40 μmol / L), ADE2-LB (SEQ ID NO.10, 5 μmol / L).

[0157] 2. dTNPs (2.5mol / L).

[0158] 3. Strand displacement DNA polymerase (8,000 units / ml).

[0159] 4. 10× reaction buffer, the composition includes: 200mM Tris-HCl, 100mM (NH 4 ) 2 SO 4 , 100mM KCl, 20mM MgSO 4 , 1% Triton X-100, pH8.8.

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Abstract

The invention relates to an LAMP (loop-mediated isothermal amplification) kit for early diagnosis of candida krusei and a special primer. The primer is designed by using an ADE2 sequence of a candida krusei strain as a target sequence. After proofing by effectivity, specificity and sensitivity experiment, the primer has the characteristics that the primer is used for the early diagnosis of candida krusei infection; the rapidity, simplicity and convenience are realized, the operability is strong, and the result judging is convenient; the problems of long diagnosis period, higher requirement on techniques of detectors and conditions of laboratory platforms, etc. in the clinical candida krusei diagnosis are solved; the primer can be prepared into the kit, and can be used for early diagnosis of candida krusei infection in clinical application.

Description

technical field [0001] The invention relates to the technical field of molecular detection, in particular to a LAMP kit for early diagnosis of Candida krusei and special primers thereof. Background technique [0002] Invasive fungal diseases (IFDs) are systemic infectious diseases caused by pathogenic fungi invading heart, liver, spleen, lung, kidney, brain, blood and other internal organs. In the past two decades, due to the widespread use of immunosuppressants, various intubation techniques and the continued spread of HIV, the morbidity and mortality of IFDs have continued to rise globally. Data from the my country Nosocomial Infection Surveillance Network shows that IFDs Now it has become the second most important component of nosocomial infection in China (accounting for about 24.1%), and yeast-like fungi dominated by Candida account for 91.4% of invasive fungal diseases. In recent years, the spectrum of pathogenic bacteria in invasive candidiasis has also undergone majo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/6895
Inventor 陈敏刘加方文捷洪南潘炜华廖万清
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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