4小结
与展望随着医学技术的不断发展，凭借人体基因密码预测相关疾病的风险性和发展进程，做到早检测、早预防、早治疗，基因诊断技术在临床的应用将有更加广阔的前景。展望未来，我们可以预见：（1）染色体水平与基因水平的检测将结合得更加紧密；（2）高通量、自动化、低成本获得更大发展；（3）PCR技术将以优化反应和拓展应用为主，随着反应速度、延伸范围和检测成本的不断优化，PCR技术将继续引领现代分子生物学的发展； （4）无创性产检将相当普及，PGD也将进入到一个更普及的阶段；（5）群体筛查的项目将更多，技术手段将更高、更快捷、更准确；（6）随着仪器成本的降低、小型化，试剂、试剂盒的大量研发，个体化、自主化检测程度将更加提高；（7）临床与基础研究结合将更加密切，研究成果将更加及时应用于临床诊防治，转化医学得到充分体现；（8）每个人一生的生老病死有望在胚胎早期就被解读、破译，疾病的预防有望得到更早期、更有效的控制；（9）可以为基因治疗寻找更多新的治疗靶点，从而促进基因治疗的迅猛发展。
参考文献：
[1]杜传书。医学遗传学（第3版）[M].北京：人民卫生出版社， 2014.
[2]傅松滨。医学遗传学（第3版）[M].北京：北京大学医学出版社， 2013.
[3]OMIM Entry Statistics. Gene description[EB/OL].
http://omim.org/statistics/entry, 2015-06-15/2015-06-17.
[4]方兴东。基 因 诊 断[EB/OL].
http://www.techcn.com.cn/index.php doc-view-113399, 2009-04-06/2015-06-17.
[5]百度百科。基因诊断[EB/OL].
http://baike.baidu.com/view/94960.htm func=retitle, 2015-03-11/2015-06-17.
[6]郭奕斌。基因诊断中测序技术的应用及优缺点[J].遗传， 2014,36（11）：1121-1130.
[7]百度百科。复杂基因[EB/OL].
http://hanyu.iciba.com/wiki/150455.shtml, 2014-05-24/2015-06-17.
[8]Liu D, Li J, Cao L, et al. Analysis of differentially ex-pressed genes in two immunologically distinct strains ofEimeria maxima using suppression subtractive hy-bridization and dot-blot hybridization[J]. Parasit Vectors,2014,7（1）：259.
[9]曹振龙，郝江叶，周艳芬，等。利用SSH方法筛选与鉴定AC3基因缺失小鼠主要嗅觉表皮内的差异表达基因[J].遗传， 2014,36（6）：574-583.
[10]Xu W, Li H, Wang R, et al. Differential expression ofgenes associated with the progression of renal disease inthe kidneys of liver-specific glucokinase gene knockoutmice[J]. Int J Mol Sci, 2013,14（3）：6467-6486.
[11]Wang R, Gao H, Xu W, et al. Differential expression ofgenes and changes in glucose metabolism in the liver ofliver-specific glucokinase gene knockout mice[J]. Gene,2013,516（2）：248-254.
[12]Ren HJ, Cui J, Yang W, et al. Identification of differen-tially expressed genes of yrichinella spiralis larvae afterexposure to host intestine milieu[J]. Plos One, 2013,8（6）：e67570.
[13]Jung J, Kim SJ, Lee KW, et al. Approaches to label-freeflexible DNA biosensors using low-temperature solution-processed In Zn O thin-film transistors [J]. Biosens Bio-electron, 2014,55:99-105.
[14]邓朝阳，宋贵生，徐军望，等。一 种 简 单 易 行 的 重 组 方法---三引物PCR法[J].科学通报， 2002,47（16）：1247-1249.
[15]朱艳，陈永井，邱玉华，等。 TP_PCR法构建抗人CD28嵌合抗体双启动子昆虫杆状病毒重组转移载体[J].生物工程学报， 2005,21（5）：832-836.
[16]Chen YX, Liu H, Zhang WB, et al. A novel tri-primerPCR method （TP-PCR） for rapid construction of fpggene [J]. Journal of Microbiological Methods, 2004, 56（3）：359-364.
[17]严昕，王均永，吴小末，等。实时荧光定量PCR检测原发性肝癌中h NTKL-BP1基因的表达[J].复旦学报（自然科学版）， 2007,3:411-416.
[18]Baker M. Digital PCR hits its stride[J]. Nature Methods,2012,9（6）：541-544.
[19]Marx V. PCR: paths to sensitivity [J]. Nature Methods,2014,11（3）：241-245.
[20]Miotke L, Lau BT, Rumma RT, et al. High sensitive de-tection and quantitation of DNA copy number and singlenucleotide variants with single colordroplet digital PCR[J]. Anal Chem, 2014,86（5）：2618-2624.
[21]Coudray-Meunier C, Fraisse A, Martin-Latil S, et al. Acomparative study of digital RT-PCR and RT-PCR forquantification of hepatitis a virus and norovirus inlettuceand water samples [J]. Int J Food Microbiol, 2015,201:17-26.
[22]Kinugasa H, Nouso K, Tanaka T, et al. Droplet digitalPCR measurement of HER2 in patients with gastric can-cer[J]. Br J Cancer, 2015,112（10）：1652-1625.
[23]Doi H, Takahara T, Minamoto T, et al. Droplet digitalpolymerase chain reaction （PCR） outperforms real-timePCR in the detection of environmental DNA from an in-vasive fish species[J]. Environ Sci Technol, 2015,49（9）：5601-5618.
[24]Zhu G, Ye X, Dong Z, et al. Highly sensitive dropletdigital PCR method for detection of EGFR-activatingmutations in plasma cell-free DNA from patients withadvanced non-amall cell lung cancer [J]. J Mol Diagn,2015,17（3）：265-272.
[25]Hayden RT, Gu Z, Sam SS, et al. Comparative evalua-tion of three commercial quantitative cytomegalovirusstandards by use of digital and real-time PCR[J]. J ClinMicrobiol, 2015,53（5）：1500-1505.
[26]Tanaka H, Yamamoto S, Nakamura A, et al. Hands-offpreparation of monodisperse emulsion droplets using apoly （dimethylsiloxane） microfluidic chip for dropletdigital PCR[J]. Anal Chem, 2015,87（8）：4134-4143.
[27]Huang H, Li S, Sun L, et al. Digital detection of multi-ple minority mutants and expression levels of multiplecolorectal cancer-related genes using digital-PCR cou-pled with bead-array[J]. Plos One, 2015,10（4）：e0123420.
[28]Floren C, Wiedemann I, Brenig B, et al. Species identi-fication and quantification in meat and products usingdroplet digital PCR （dd PCR） [J]. Food Chem, 2015,15;173:1054-1058.
[29]Devonshire AS, Honeyborne I, Gutteridge A, et al.Highly reproducible absolute quantification of mycobac-terim tuberculosis complex by digital PCR [J]. AnalChem, 2015,87（7）：3706-3713.
[30]Zhang BO, Xu CW, Shao Y, et al. Comparison ofdroplet digital PCR and conventional quantitative PCRfor measuring EGFR gene mutation [J]. Exp Ther Med,2015, 9（4）：1383-1388.
[31]Kinz E, Leiherer A, Lang AH, et al. Accurate quantita-tion of JAK2 V617F allele burden by array-based digitalPCR[J]. Int J Lab Hematol, 2015, 37（2）：217-224.
[32]Doi H, Uchii K, Takahara T, et al. Use of dropet digitalPCR for estimation of fish abundance and biomass inenviromental DNA surveys [J]. Plos One, 2015,10（3）：e0122763.
[33]Miyake K, Yang C, Minakuchi Y, et al. Comparison ofgenomic and epigenomic expression in monozygotictwins discordant for rett syndrome[J]. Plos One, 2013, 8（6）：e66729.
[34]Liu W, Hu T, Chen Y, et al. Development and valida-tion of a tetra-primer amplification refractory mutationaystem-polymerase chain reaction combined with melt-ing analysis-assay for clinical JAK2 V617F mutation de-tection[J]. Mol Diagn Ther, 2014,18（5）：579-585.
[35]Cheng C, Zhou Y, Yang C, et al. Detection of rare pointmutation via allele-specific amplification in emulsionPCR[J]. BMB reports, 2013,46（5）：270-275.
[36]李荣，毕博文，沈晓婷，等。软骨发育不全植入前遗传学诊断的方法学研究[J]分子诊断与治疗杂志， 2014,6（6）：372-377.
[37]Liu W, Smith DI, Rechtzigel KJ, et al. Denaturing highperformance liquid chromatography （DHPLC） used inthe detection of germline and somatic mutations[J]. Nu-cleic Acids Res, 1998,26（6）：1396-1400.
[38]新浪博客。 DHPLC变性高效液相色谱原理[EB/OL].http://blog.sina.com.cn/s/blog_4d82e72b01000axl.html.2007-07-22/2015-06-17.
[39]Reed GH, Kent JO, Wittwer CT. High-resolution DNAmelting analysis for simple and efficient molecular diag-nostics[J]. Pharmacogenomics, 2007,8（6）：597-608.
[40]Schouten JP, Mc Elgunn CJ, Waaijer R, et al. Relativequantification of 40 nucleic acid sequences by multiplexligation-dependent probe amplification[J]. Nucleic AcidsRes, 2002,30（12）：e57.
[41]胡晓，李汶，卢光琇。多重连接依赖性探针扩增技术及其应用进展。现代生物医学进展[J]. 2010,10（2）：362-365.
[42]Necker J, Kovac M, Attenhofer M, et al. Detection ofAPC germ line mosaicism in patients with de novo fa-milial adenomatous polyposis: a plea for the proteintruncation test[J]. J Med Genet, 2011,48（8）：526-529.
[43]新浪博客。瞬时基因表达技术[EB/OL].
http://blog.sina.com.cn/s/blog_4e0c3abb010096ph.html, 2008 -05 -09/2015-06-17.
[44]中华论文网。概述基因芯片技术原理等[EB/OL].
http://www.zhonghualw.com/xumushouyi/net201202090638543029.html, 2012-02-09/2015-06-17.
[45]好搜问答。基因芯片技术[EB/OL].
http://wenda.so.com/q/1378647361064338 src=140, 2013-09-08/2015-06-17.
[46]滕牧洲，马文丽。走进医学研究领域中的基因芯片技术[J].分子诊断与治疗杂志， 2014,6（6）：361-365.
