Evaluation method for identifying regional applicability of specific marker

Abstract

The invention discloses an evaluation method for identifying the applicability of a specific marker region, which comprises the following steps: S1, collecting a sample, and extracting DNA; s2, selecting a specific primer; s3, making a qPCR reaction and a standard curve; s4, selecting sensitivity, specificity, source strength characteristics, a 25th / 75th threshold value and a target source 25th / 75th as indexes for evaluating the applicability of the primer, and calculating the value of each index; and S5, performing weighting on five types of parameter indexes including primer sensitivity, specificity, source strength characteristics, 25th / 75th threshold values and target source 25th / 75th by utilizing an analytic hierarchy process to obtain comprehensive adaptability evaluation of each primer. According to the method disclosed by the invention, the host specific marker which has the advantages of specificity, relatively high detection level and relatively stable detection concentrationcan be effectively screened out and is used for subsequent water body microbial pollution quantitative source analysis work.

Description

technical field [0001] The invention relates to the research field of microbial pollution prevention and control, in particular to an evaluation method for regional applicability of identification specific markers. Background technique [0002] Human and animal feces contain a variety of pathogenic microorganisms and enteroviruses. Discharge of untreated (or incompletely treated) feces into natural water bodies not only causes serious water microbial pollution, but also brings great harm to human health. big risk. For example, there are reports of norovirus gastroenteritis cases in many regions of China every year, and the main route of transmission is "fecal-oral" transmission. The novel coronavirus (2019-nCoV) outbreak that broke out at the end of 2019 also has the possibility of "fecal-oral" transmission. Therefore, how to quickly and accurately identify the source of microbial contamination and evaluate the contribution rate of each source has become a key link to effe...

AI Technical Summary

Problems solved by technology

In the past, two indexes, sensitivity and specificity, were usually used to evaluate the applicability of primers. However, these two types of evaluation factors can only conduct qualitative analysis on the characteristics of primers, and it is difficult to reflect the expression levels of primers in different regions. Unable to quantify microbial contamination from different host sources due to differences in detection concentrations of each primer

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