微流控芯片核酸分析系统及其精准医学应用 
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Microfluidic Chip Based Nucleic Acid Analyzer and Its Application in Precision Medicine
1 清华大学医学院， 北京 100084
2 北京航空航天大学北京学院， 北京100191
3 首都医科大学生物医学工程学院， 北京 100069
4 生物芯片北京国家工程研究中心， 北京 102206
图 & 表
图 1. 核酸恒温扩增原理示意图。(a)启动阶段;(b)循环阶段
Fig. 1. Principle diagram of isothermal nucleic acid amplification. (a) Initialization phase; (b) circulatory phase
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图 2. 24通道微流控芯片设计结构示意图
Fig. 2. Structural diagram of microfluidic chip with 24 channels
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图 3. 核酸分析仪原理结构
Fig. 3. Schematic diagram of nucleic acid analyzer
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图 4. (a)(b)微流控芯片核酸分析仪的实物照片;(c)微流控芯片试剂盒照片
Fig. 4. (a)(b) Photos of micro-fluidic chip based nucleic acid analyzer; (c) photo of micro-fluidic chip kit
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图 5. 微流控芯片核酸恒温扩增检测结果。(a)灵敏度测试分析结果;(b)线性度测试结果;(c)特异性测试结果
Fig. 5. Results of microfluidic chip based isothermal nucleic acid amplification. (a) Result of sensitivity test; (b) result of linearity test; (c) result of specificity test
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表 1病原菌的特异基因探针设计举例
Table1. Example for gene-specific probes of pathogenic bacteria
| Testing index | Probe code | Base sequence of probe |
|---|
| Mycoplasmapneumonia | F3 | CTCACCGTAGTGGGACA | | B3 | GCCCCGGGATTTTCACC | | FIP | CGTCAGGGCGGGTGTAGCTCTTCACAAGTACCACCACGAC | | BIP | TGCGCCACACCAATGCCATGGGAGGGAGGAAAAGCT | | LF | ATTGCTGGCGCTTGAGC | | LB | CGCGCTTAACCCCGTGA |
|
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表 2临床样品检测实验结果
Table2. Results of clinical sample test
| Test index | Result fromdevelopedanalyzer | Result from ABI 7500 | Positivecoincidencerate /% | Negativecoincidencerate /% | Totalcoincidencerate /% |
|---|
| | | PositiveNo. | NegativeNo. | SubtotalNo. |
|---|
| Positive No. | 23 | 1 | 24 | | | | | Sau | Negative No. | 0 | 76 | 76 | 100 | 98.7 | 99.0 | | Subtotal No. | 23 | 77 | 100 | | | | | Positive No. | 22 | 1 | 23 | | | | | MRSA | Negative No. | 0 | 77 | 77 | 100 | 98.7 | 99.0 | | Subtotal No. | 22 | 78 | 100 | | | | | Positive No. | 14 | 1 | 15 | | | | | Mpn | Negative No. | 1 | 84 | 85 | 93.3 | 98.8 | 98.0 | | Subtotal No. | 15 | 85 | 100 | | | | | Positive No. | 59 | 3 | 62 | | | | | Total | Negative No. | 1 | 237 | 238 | 98.3 | 98.8 | 98.7 | | Subtotal No. | 60 | 240 | 300 | | | |
|
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表 3测试结果不一致样品分析
Table3. Analysis of discrepancy samples of test result
| Sample No. | ABI 7500 | Developedanalyzer | Reanalysisby ABI 7500 | Reanalysis bydeveloped analyzer | Sequencing |
|---|
| 22 | Mpn | Sau | Mpn | Sau | Sau | | 32 | Sau | Sau, MRSA | Sau | Sau, MRSA | Sau, MRSA |
|
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黄世光, 靳翔宇, 林荣赞, 林雪, 薛宁, 范云倩, 祖国, 马丽, 罗贤波, 黄国亮. 微流控芯片核酸分析系统及其精准医学应用[J]. 中国激光, 2018, 45(3): 0307002. Huang Shiguang, Jin Xiangyu, Lin Rongzan, Lin Xue, Xue Ning, Fan Yunqian, Zu Guo, Ma Li, Luo Xianbo, Huang Guoliang. Microfluidic Chip Based Nucleic Acid Analyzer and Its Application in Precision Medicine[J]. Chinese Journal of Lasers, 2018, 45(3): 0307002.
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