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
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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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