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Characteristic sequence identification method of common toxigenic fungi in drug quality control

A characteristic sequence and drug quality technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of complex types of characteristic sequences, and there is no method for identifying the characteristic sequences of toxin-producing fungal systems. Stable and reliable, improved identification level, strong discrimination effect

Pending Publication Date: 2020-03-13
SHANGHAI INST FOR FOOD & DRUG CONTROL
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Problems solved by technology

[0004] The research on the identification method of the existing characteristic sequence is mainly to identify a certain genus or a specific type of fungi. There are many and complicated types of characteristic sequences, and there is no systematic characteristic sequence identification method for common toxin-producing fungi in the pharmaceutical field.

Method used

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  • Characteristic sequence identification method of common toxigenic fungi in drug quality control
  • Characteristic sequence identification method of common toxigenic fungi in drug quality control
  • Characteristic sequence identification method of common toxigenic fungi in drug quality control

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Embodiment 1

[0041] The present invention provides a characteristic sequence identification method for measuring more than 30 kinds of fungi belonging to 8 genera, wherein the types of fungi used are as shown in Table 1, and the specific identification steps are as follows:

[0042] Step 1. Isolation and purification of bacterial strains

[0043] Invert the appropriate solid medium plate, use the inoculation loop to take the bacteria to be tested or its test solution, inoculate 1 to 3 points upward, and then place the plate upright for culture.

[0044] Step 2. Nucleic acid extraction

[0045]Take about 20 mg of bacteria and place them in a centrifuge tube, grind, then add 400 μl of CTAB buffer (50mmol / LTris-HCl pH8, 0.7mol / L NaCl, 10mmol / L EDTA pH8, 2% CTAB) and 4μl 2-mercaptoethanol; Water bath at 65°C for 1 hour, shake once every 10 minutes during the water bath; add 200 μl saturated phenol-chloroform-isoamyl alcohol (25:24:1, v / v / v) solution, mix thoroughly by inverting, and store in ...

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Abstract

The invention discloses a characteristic sequence identification method of common toxigenic fungi in drug quality control. The characteristic sequences used are ITS and LSU, and their amplification primer pair sequences are shown as SEQ ID NO.1 to SEQ ID NO.2 and SEQ ID NO.3 to SEQ ID NO.4, respectively. The characteristic sequences adopted by the invention have a moderate length and meet the length requirements of characteristic sequence, have good universality and can be applied to common toxigenic fungi, have strong discrimination and can effectively distinguish the sequence differences offungi at the level of ''genus'' or ''species'', have high accuracy, can be used for fungus identification separately, and can be used to further improve the identification level of strains in the wayof combining the two characteristic sequences, and the results are stable and reliable. In conclusion, the characteristic sequence identification method of common toxigenic fungi in the invention provides an effective and widely applicable fungus identification method for drug quality control.

Description

technical field [0001] The invention relates to the field of fungal identification, in particular to a characteristic sequence identification method of common toxin-producing fungi in drug quality control. Background technique [0002] With the rapid development of the pharmaceutical industry, incidents of microbial contamination of related products are also increasing. Contaminating microorganisms include not only bacteria but also fungi, some of which can also produce highly toxic mycotoxins. The quality of medicines and the safety of patients' medicines pose a great risk. In order to control the quality of drugs and avoid harm to the health of patients, "Chinese Pharmacopoeia" (2015 edition), "United States Pharmacopoeia" (USP 41), "European Pharmacopoeia" (EP 9.0) and "Japanese Pharmacopoeia" (JP 17) require Check for pathogenic bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in drugs, and clarify that in addition to the pathogenic b...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6806C12Q1/04
CPCC12Q1/6895C12Q1/6806Y02A50/30
Inventor 蒋波冯震杨美成刘浩秦峰缪天瑶
Owner SHANGHAI INST FOR FOOD & DRUG CONTROL
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