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Method for discriminating microorganism by utilizing Fourier infrared spectrum

A technology of infrared spectroscopy and microorganisms, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc., can solve problems such as difficult general laboratory promotion, and achieve rapid identification, strong specificity, and low cost low effect

Active Publication Date: 2009-10-14
NAT INST FOR FOOD & DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have good specificity and high sensitivity, but they need professionals to complete and are difficult to promote in ordinary laboratories

Method used

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  • Method for discriminating microorganism by utilizing Fourier infrared spectrum
  • Method for discriminating microorganism by utilizing Fourier infrared spectrum
  • Method for discriminating microorganism by utilizing Fourier infrared spectrum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Step 1 Selection and cultivation of control microorganisms

[0087] In order to identify the six control bacteria specified in the Chinese Pharmacopoeia, a total of 191 strains of bacteria belonging to 9 genera and 23 species in 4 families were selected as control microorganisms to construct a reference spectrum library. See Table 1 for specific strains and quantities.

[0088] Table 1

[0089]

[0090]

[0091] Step 2 Spectral determination of control microorganisms

[0092] The above-mentioned control microorganisms were purified on a TSA medium (BD Company, USA) plate to form a single colony, and then a single colony was selected to be streaked and inoculated on the plate, and cultured at 35°C for 24±2h.

[0093] Use a calibrated vibrating platinum inoculation loop with a diameter of 1 mm (product of Braun, Germany) to take 1-2 rings of bacterial lawn from the place where the colonies converge on the TSA medium, place them in 100 μl of sterilized water, and s...

Embodiment 2

[0116] The test was carried out using the same method and steps as in Example 1, except that the microorganisms to be tested were combined using two identification models to improve the correct rate of identification.

[0117] The specific procedures and results judgment methods are as follows: Figure 5 shown.

[0118] The original spectrum and the average spectrum of the strains identified correctly in the first step were brought into the second identification method respectively, and the correct identification rate was 92.2% (379 / 411). The strains and results of validation errors are shown in Table 4. It can be seen from Table 4 that identification errors mainly occurred when E. coli was identified as other Enterobacteriaceae microorganisms, that is, false negative results were presented.

[0119] In view of the false negative results caused by the original qualitative analysis identification model, the above spectra that were not correctly identified were added to the mo...

Embodiment 3

[0124] Embodiment 3 differentiates under species

[0125] To distinguish bacterial species below species, such as serotype level. In this embodiment, a series of Salmonella were selected, and their serotypes and IR spectra were determined for cluster analysis. see details Image 6 , where the serum types include O9, O4, and O7, the selected spectral bands are 1202-899, 901-698, and the reproducibility value is 1. The spectrum processing method is First derivative+Vector normalization, and the algorithm is Ward’s algorithm. As can be seen from the figure, the IR patterns are well classified by serotype.

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Abstract

The invention relates to a method for discriminating microorganism by utilizing Fourier infrared spectrum, comprising the following steps of: a) culturing control microorganism; b) collecting infraredspectrum of the control microorganism; c) establishing a discriminating model of the microorganism in one or a plurality of spectral sections within the interval between 3000-2800cm<-1> and 1800-700c m<-1>; d) culturing the microorganism to be detected under the same condition with the step a); e) collecting the infrared spectrum of the microorganism to be detected under the same condition with the step b); and f) substituting the infrared spectrum obtained in the step e) into the discriminating model of the microorganism and determining the attribution of the microorganism to be detected. Thediscriminating model of the microorganism can realize fast discrimination and does not need to carry out pre-selection to the classification of the microorganism to be detected by using other taxonom ic methods; the method has strong specificity, can carry out classification according to the units of families, classes, species and the like, and even can discriminate to the level of subspecies and serological types; and once the discriminating model is established, the cost for discriminating microorganism is extremely low.

Description

technical field [0001] The present invention relates to a method for identifying microorganisms using Fourier transform infrared spectroscopy. More specifically, it relates to a method for establishing a microorganism identification model using Fourier transform infrared spectroscopy and using the model to identify microorganisms, such as contaminating bacteria, in medicines. Background technique [0002] The detection of microorganisms in medicines is an important item in the quality inspection of medicines, and the accuracy of the test results directly determines its credibility. Among them, the inspection of microbial contamination should first confirm whether there are pathogenic microorganisms. In many cases, further identification of the pathogenic microorganisms present is required. [0003] Under normal circumstances, pharmacopoeias, such as the Chinese Pharmacopoeia, require microbial limit inspection (including the determination of the number of contaminated bact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/35C12Q1/04G01N21/3563G01N21/3577
Inventor 胡昌勤裴琳马仕洪戴翚肖璜
Owner NAT INST FOR FOOD & DRUG CONTROL
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