[논문]수질분석에 사용되는 qPCR기술

김원재; 황윤정; 이민혜; 정민섭

doi:10.14478/ace.2022.1027

수질분석에 사용되는 qPCR기술
Utilization of qPCR Technology in Water Treatment 원문보기

공업화학 = Applied chemistry for engineering, v.33 no.3, 2022년, pp.235 - 241

김원재 (홍익대학교 화학공학과) , 황윤정 (홍익대학교 화학공학과) , 이민혜 (홍익대학교 화학공학과) , 정민섭 (홍익대학교 화학공학과)

초록
AI-Helper

유엔이 발표한 세계 물개발 보고서는 2030년까지 식수가 현재보다 40% 감소할 것으로 전망하고 있다. 이는 물의 양이 감소하는 것이 아니라, 환경오염으로 인해 상수원이 오염되는 것을 말한다. 미생물이 수질에 깊은 연관이 있기 때문에 미생물의 분석은 수질관리에 매우 중요하다. 현재 미생물 분석에 사용되는 방법은 배양 후 현미경을 통한 모양과 형태를 분석하는 것이 가장 일반적이나, 유전자분석 기술이 발달함에 따라 현미경을 통한 미생물 분석 방식에 qPCR(quantitative polymerase chain reaction) 적용이 가능해졌고 활용방법 등이 연구되었다. 그 중에는 역전사 단계를 추가하여 RNA 분석에 용이성을 부여한 RT-qPCR법과 미생물 배양분석에 접목시켜 검사시간을 단축시키는 ICC-qPCR, 자연에서 채취한 샘플의 위양성율을 감소시키는 데 용이한 viability qPCR, 다중분석에 용이한 multiplex qPCR, 소량의 샘플만으로 분석이 가능한 microfluidic qPCR법 등이 있다. 본 논문에서는 이처럼 qPCR 방법이 미생물 분석에 적용되는 사례와 방식의 원리, 그리고 발전 방향에 대해 소개하고자 한다.

Abstract ▼ AI-Helper

According to the World Water Development Report 2015 released by the United Nations, drinking water is expected to decrease by 40% by 2030. This does not mean that the amount of water decreases, but rather that the water source is contaminated due to environmental pollution. Because microbes are deeply related to water quality, the analysis of microbe is very important for water quality management. While the most common method currently used for microbial analysis is microscopic examination of the shape and feature after cell culture, as the gene analysis technology advances, quantitative polymerase chain reaction (qPCR) can be applied to the microscopic microbiological analysis, and the application method has been studied. Among them, a reverse transcription (RT) step enables the analysis of RNA by RT-PCR. Integrated cell culture (ICC)-qPCR shortens the test time by using it with microbial culture analysis, and viability qPCR can reduce the false positive errors of samples collected from natural water source. Multiplex qPCR for improved throughput, and microfluidic qPCR for analysis with limited amount of sample has been developed In this paper, we introduce the case, principle and development direction of the qPCR method applied to the analysis of microorganisms.

주제어

표/그림 (5)

그림 Figure 1. (A) qPCR with DNA intercalating dyes. The reporter dyes become fluorescent as inserted to double strand DNA selectively during extension step. (B) The principle of TaqMan probes. Fluorescence occurs as the reporter dye separates from the quencher dye in extension step. (C) The amplification plots from the extension phase fluorescent emission data. Ct values are calculated by determining the point at which the fluorescence exceeds a threshold limit. (D) Calibration curve showing Ct changes according to template DNA concentration.
그림 Figure 2. Schematics of the RT-qPCR principle and process.
그림 Figure 3. The method for confirming the cytopathic effect requires a culture period of 5 to 7 days, whereas the method for qPCR requires less than 24 hours of incubation time.
그림 Figure 4. Schematics of the viability qPCR. The viability dye can enter the damaged cells through the cell membrane and be bound to DNA when irradiated with UV light.
그림 Figure 5. Schematic of the microfluidic qPCR operation. (A) The PCR mixture loading step, (B) The crosssectional view of the PCR array chip with a localized thermal electric cooler (TEC) and heat-sink. (C) Sample loading and thermal cycling. (D) Photograph of the PDMS-glass chip comprising an array of nine microreactors[22].

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표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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