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Application of Grading of Recommendations Assessment, Development, and Evaluation (GRADE) to the guideline

Hong Zhao, Fengxia Liang, Yigong Fang, Baoyan Liu

Frontiers of Medicine 2017, Volume 11, Issue 4, Pages 590-594 doi: 10.1007/s11684-017-0537-4

Abstract: Grading of Recommendations Assessment, Development, and Evaluation (GRADE) offers a worldwide approach to guidelineIn this study, we review the advantages and limitations of the GRADE approach in the guideline developmentIn the guideline development for clinical practice with acupuncture and moxibustion, we suggest that

Keywords： guideline acupuncture GRADE

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Design and use of group-specific primers and probes for real-time quantitative PCR

Juntaek LIM, Seung Gu SHIN, Seungyong LEE, Seokhwan HWANG

Frontiers of Environmental Science & Engineering 2011, Volume 5, Issue 1, Pages 28-39 doi: 10.1007/s11783-011-0302-x

Abstract: Real-time quantitative polymerase chain reaction (qPCR) has gained popularity as a technique to detect and quantify a specific group of target microorganisms from various environmental samples including soil, water, sediments, and sludge. Although qPCR is a very useful technique for nucleic acid quantification, accurately quantifying the target microbial group strongly depends on the quality of the primer and probe used. Many aspects of conducting qPCR assays have become increasingly routine and automated; however, one of the most important aspects, designing and selecting primer and probe sets, is often a somewhat arcane process. In many cases, failed or non-specific amplification can be attributed to improperly designed primer-probe sets. This paper is intended to provide guidelines and general principles for designing group-specific primers and probes for qPCR assays. We demonstrate the effectiveness of these guidelines by reviewing the use of qPCR to study anaerobic processes and biologic nutrient removal processes. qPCR assays using group-specific primers and probes designed with this method, have been used to successfully quantify 16S ribosomal Ribonucleic Acid (16S rRNA) gene copy numbers from target methanogenic and ammonia- oxidizing bacteria in various laboratory- and full-scale biologic processes. Researchers with a good command of primer and probe design can use qPCR as a valuable tool to study biodiversity and to develop more efficient control strategies for biologic processes.

Keywords： absolute quantification design guideline primer probe real-time quantitative polymerase chain reaction