{"id":5412,"date":"2022-04-12T08:08:04","date_gmt":"2022-04-12T00:38:04","guid":{"rendered":"https:\/\/abtvn.com\/?p=5412"},"modified":"2023-09-19T17:01:47","modified_gmt":"2023-09-19T09:31:47","slug":"ky-thuat-xet-nghiem-pcr-va-ung-dung-trong-chuan-doan-sinh-hoc-phan-tu","status":"publish","type":"post","link":"https:\/\/abtvn.com\/en\/ky-thuat-xet-nghiem-pcr-va-ung-dung-trong-chuan-doan-sinh-hoc-phan-tu\/","title":{"rendered":"K\u1ef9 thu\u1eadt PCR v\u00e0 \u1ee9ng d\u1ee5ng trong ch\u1ea9n \u0111o\u00e1n sinh h\u1ecdc ph\u00e2n t\u1eed<\/strong>"},"content":{"rendered":"

PCR<\/strong> is a technique that has been mentioned a lot recently in medical diagnostic testing, especially in diagnostic testing for covid-19. PCR testing method<\/strong> is used in the laboratory for the purpose of cloning millions of copies of a target DNA fragment. So what is the main working principle of PCR? And what is the main application of PCR test in medical diagnosis?<\/p>\n\n\n\n

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What is PCR<\/strong><\/h2>\n\n\n\n

PCR (Polymerase Chain Reaction)<\/strong> <\/a>is a fast and efficient technique used to amplify a small target DNA fragment that produces millions of copies of DNA. In other words, PCR makes it possible to make millions of copies of a particular DNA sequence from a small initial sample or even a single copy.<\/p>\n\n\n\n

PCR<\/strong> is considered an important process for a wide range of genetic technologies in medical and biological research. Especially in molecular biology tests, PCR is an indispensable process to help people study deeply into the analysis of DNA\/RNA genetic material.  <\/p>\n\n\n\n

PCR principle<\/strong><\/h2>\n\n\n\n
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\"Nguy\u00ean
Principle of PCR<\/em><\/figcaption><\/figure>\n\n\n\n

Ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n d\u1ef1a tr\u00ean nguy\u00ean t\u1eafc khu\u1ebfch \u0111\u1ea1i m\u1ed9t \u0111o\u1ea1n tr\u00ecnh t\u1ef1 DNA \u0111\u1eb7c hi\u1ec7u in vitro do s\u1ef1 x\u00fac t\u00e1c c\u1ee7a enzyme DNA polymerase. S\u1ef1 khu\u1ebfch \u0111\u1ea1i n\u00e0y \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n nh\u1edd c\u00e1c chu tr\u00ecnh lu\u00e2n nhi\u1ec7t (c\u00f3 th\u1ec3 20-40 chu k\u1ef3) g\u1ed3m 3 b\u01b0\u1edbc:<\/p>\n\n\n\n

+ Denaturation 95\u20700<\/sup>C: thermal separation of double-stranded DNA into single-stranded DNA.<\/p>\n\n\n\n

+ Annealing 40\u2070C0<\/sup>- 70\u2070C:0<\/sup>allows primers to be paired with the DNA template.<\/p>\n\n\n\n

+ Elongation 72\u2070C:0<\/sup>new strand is synthesized by the principle of complementarity by DNA polymerase<\/p>\n\n\n\n

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C\u00e1c th\u00e0nh ph\u1ea7n c\u1ea7n thi\u1ebft \u0111\u1ec3 th\u1ef1c hi\u1ec7n ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i<\/strong><\/h2>\n\n\n\n
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\"C\u00e1c
Basic components in a PCR reaction<\/em><\/figcaption><\/figure>\n\n\n\n

When setting up a PCR reaction, regardless of the DNA sample, the following components are required:<\/p>\n\n\n\n

 - DNA template<\/p>\n\n\n\n

– M\u1ed3i (Primer): L\u00e0 tr\u00ecnh t\u1ef1 oligonucleotide c\u00f3 kh\u1ea3 n\u0103ng b\u1eaft c\u1eb7p chuy\u00ean bi\u1ec7t v\u1edbi \u0111\u1ea7u 5\u2019 hay \u0111\u1ea7u 3\u2019 c\u1ee7a \u0111o\u1ea1n DNA c\u1ea7n nh\u00e2n b\u1ea3n sao. M\u1ed3i \u0111\u01b0\u1ee3c d\u00f9ng theo t\u1eebng c\u1eb7p m\u1ed3i g\u1ed3m m\u1ed3i xu\u00f4i v\u00e0 m\u1ed3i ng\u01b0\u1ee3c t\u00ednh theo chi\u1ec1u phi\u00ean m\u00e3. M\u1ed3i xu\u00f4i c\u00f3 tr\u00ecnh t\u1ef1 nucleotide c\u00f9ng chi\u1ec1u v\u1edbi s\u1ee3i mang m\u00e3, m\u1ed3i ng\u01b0\u1ee3c c\u00f3 tr\u00ecnh t\u1ef1 nucleotide b\u1ed5 sung v\u1edbi s\u1ee3i mang m\u00e3 c\u1ee7a \u0111o\u1ea1n DNA c\u1ea7n nh\u00e2n b\u1ea3n sao.<\/p>\n\n\n\n

- dNTP (deoxy nucleotide triphosphate): dNTP is a mixture of 4 components needed for DNA synthesis: dATP, dGTP, dCTP, dTTP.<\/p>\n\n\n\n

- DNA polymerase: an enzyme that prolongs the new strand that is complementary to the template strand from the primers with heat-resistant characteristics and does not lose its activity at high temperatures of the denaturation step.<\/p>\n\n\n\n

– Dung d\u1ecbch \u0111\u1ec7m ( buffer): th\u00e0nh ph\u1ea7n Tris-HCl pH 8,3 ; KCl, MgCl2 v\u00e0 Gelatin. Dung d\u1ecbch \u0111\u1ec7m c\u00f3 vai tr\u00f2 trong vi\u1ec7c t\u1ea1o s\u1ef1 k\u1ebft h\u1ee3p ch\u1eb7t ch\u1ebd gi\u1eefa c\u00e1c dNTP, k\u00edch ho\u1ea1t v\u1ecb tr\u00ed ho\u1ea1t \u0111\u1ed9ng c\u1ee7a DNA polymerase. Dung d\u1ecbch \u0111\u1ec7m c\u00f3 \u1ea3nh h\u01b0\u1edfng m\u1ea1nh m\u1ebd \u0111\u1ebfn t\u00ednh \u0111\u1eb7c hi\u1ec7u v\u00e0 hi\u1ec7u su\u1ea5t c\u1ee7a ph\u1ea3n \u1ee9ng PCR .<\/p>\n\n\n\n

A complete PCR reaction can be performed in a few hours, or even less than an hour with certain high-speed machines.<\/p>\n\n\n\n

Sau khi ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i \u0111\u01b0\u1ee3c ho\u00e0n th\u00e0nh trong m\u00e1y lu\u00e2n nhi\u1ec7t, m\u1ed9t ph\u01b0\u01a1ng ph\u00e1p g\u1ecdi l\u00e0 \u0111i\u1ec7n di tr\u00ean gel agarose c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 ki\u1ec3m tra s\u1ed1 l\u01b0\u1ee3ng v\u00e0 k\u00edch th\u01b0\u1edbc c\u1ee7a c\u00e1c \u0111o\u1ea1n DNA \u0111\u01b0\u1ee3c t\u1ea1o ra.<\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n
\"K\u1ebft
Image of PCR product sample loading (A) and electrophoresis results (B) on agarose gel stained with Ethidium Bromide <\/em><\/figcaption><\/figure>\n\n\n\n
<\/div>\n\n\n\n

Application of PCR in medical field<\/strong><\/h2>\n\n\n\n

K\u1ef9 thu\u1eadt x\u00e9t nghi\u1ec7m PCR<\/strong> l\u00e0 m\u1ed9t c\u00f4ng c\u1ee5 ch\u1ea9n \u0111o\u00e1n tuy\u1ec7t v\u1eddicho ph\u00e9p con ng\u01b0\u1eddi nh\u00e2n l\u00ean c\u1ea3 tri\u1ec7u \u0111o\u1ea1n DNA (RNA) trong th\u1eddi gian r\u1ea5t ng\u1eafn v\u1edbi \u0111\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u cao. \u1ee8ng d\u1ee5ng trong ch\u1ea9n \u0111o\u00e1n y h\u1ecdc bao g\u1ed3m:<\/p>\n\n\n\n

+ Diagnosis of human infectious diseases<\/strong>: Based on detection of infectious bacteria or viruses such as HIV, tuberculosis (TB), Hepatitis B (HBV), Hepatitis C (HCV) Cervical cancer (HPV)...<\/p>\n\n\n\n

+ Ch\u1ea9n \u0111o\u00e1n c\u00e1c b\u1ec7nh di truy\u1ec1n, S\u1eeda \u0111\u1ed5i c\u00e1c \u0111o\u1ea1n DNA.<\/p>\n\n\n\n

+ Forensic analysis: Identification of genetic fingerprints, DNA testing, blood relations, criminal investigation..<\/p>\n\n\n\n

+ DNA cloning: Allows researchers to make identical copies of a piece of DNA, studying gene expression<\/p>\n\n\n\n

In addition to the medical field, the PCR method is also applied in the production of pathogen diagnostic kits in a variety of other fields, including: Human disease identification, veterinary medicine, aquaculture, food microbiology, etc. identification,..<\/p>\n\n\n\n

Hi\u1ec7n nay, c\u00f4ng ty ABT<\/strong> l\u00e0 1 doanh nghi\u1ec7p h\u00e0ng \u0111\u1ea7u \u00e1p d\u1ee5ng ph\u01b0\u01a1ng ph\u00e1p PCR, Real-time PCR th\u00e0nh c\u00f4ng trong vi\u1ec7c s\u1ea3n xu\u1ea5t c\u00e1c b\u1ed9 kit ch\u1ea9n \u0111o\u00e1n m\u1ea7m b\u1ec7nh. \u0110\u1ec3 bi\u1ebft th\u00eam th\u00f4ng tin chi ti\u1ebft v\u1ec1 k\u1ef9 thu\u1eadt x\u00e9t nghi\u1ec7m n\u00e0y, c\u0169ng nh\u01b0 kh\u00e1m ph\u00e1 c\u00e1c b\u1ed9 kit ch\u1ea9n \u0111o\u00e1n c\u1ee7a ABT trong Y h\u1ecdc<\/strong>: <\/p>\n\n\n\n