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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front Med    2012, Vol. 6 Issue (2) : 173-186     DOI: 10.1007/s11684-012-0195-5
REVIEW |
Nucleic acid-based diagnostics for infectious diseases in public health affairs
Albert Cheung-Hoi Yu1,2(), Greg Vatcher2, Xin Yue2, Yan Dong2, Mao Hua Li2, Patrick H. K. Tam2, Parker Y.L. Tsang2, April K.Y. Wong2, Michael H.K. Hui2, Bin Yang2, Hao Tang2, Lok-Ting Lau2()
1. Neuroscience Research Institute; Department of Neurobiology, School of Basic Medical Sciences; Key Laboratory for Neuroscience, Ministry of Education; Key Laboratory for Neuroscience, Ministry of Health; Peking University, Beijing 100191, China; 2. Hai Kang Life Corporation Limited, Hong Kong SAR, China and Beijing 100176, China
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Abstract  

Infectious diseases, mostly caused by bacteria and viruses but also a result of fungal and parasitic infection, have been one of the most important public health concerns throughout human history. The first step in combating these pathogens is to get a timely and accurate diagnosis at an affordable cost. Many kinds of diagnostics have been developed, such as pathogen culture, biochemical tests and serological tests, to help detect and fight against the causative agents of diseases. However, these diagnostic tests are generally unsatisfactory because they are not particularly sensitive and specific and are unable to deliver speedy results. Nucleic acid-based diagnostics, detecting pathogens through the identification of their genomic sequences, have shown promise to overcome the above limitations and become more widely adopted in clinical tests. Here we review some of the most popular nucleic acid-based diagnostics and focus on their adaptability and applicability to routine clinical usage. We also compare and contrast the characteristics of different types of nucleic acid-based diagnostics.

Keywords nucleic acid-based diagnostics      infectious disease      PCR      NASBA      LAMP      microarray      LOAC      public health affairs     
Corresponding Authors: Yu Albert Cheung-Hoi,Email:achy@haikanglife.com, achy@hsc.pku.edu.cn; Lau Lok-Ting,Email:terence.lau@haikanglife.com   
Issue Date: 05 June 2012
URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-012-0195-5     OR     http://academic.hep.com.cn/fmd/EN/Y2012/V6/I2/173
Fig.1  Enhanced real-time PCR (ERT-PCR). The template sequence (RNA or DNA) is amplified with a set of specific primers. The PCR product is then used as a template for a second round of amplification using a second set of nested primers. During this second round of PCR a detection probe is included to quantitate the reaction in real-time. The probe contains a fluorescent reporter (R) and quencher (Q). When the reporter and quencher are close together there is no fluorescent signal. As the PCR continues the probe is displaced and cleaved, releasing the reporter which emits a detectable fluorescent signal.
Fig.2  Nucleic acid sequence-based amplification (NASBA). An RNA sequence of interest is amplified with reverse transcriptase and a sequence-specific primer. The DNA/RNA hybrid is then treated with RNase-H to hydrolyze the RNA. Reverse transcriptase and a second primer are then used to generate a double stranded DNA with incorporated RNA polymerase (RNAP) and ECL (or EOC) binding sites (see text for details). RNAP produces an RNA template of the target sequence which is used to repeat the whole cycle, allowing large amounts of the target sequence to be generated and detected.
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