{"id":7479,"date":"2023-01-04T09:57:38","date_gmt":"2023-01-04T02:27:38","guid":{"rendered":"https:\/\/abtvn.com\/?p=7479"},"modified":"2023-04-06T14:41:11","modified_gmt":"2023-04-06T07:11:11","slug":"ky-thuat-khuech-dai-sinh-hoc-phan-tu-nested-pcr","status":"publish","type":"post","link":"https:\/\/abtvn.com\/en\/ky-thuat-khuech-dai-sinh-hoc-phan-tu-nested-pcr\/","title":{"rendered":"Nested PCR – K\u1ef9 thu\u1eadt PCR n\u00e2ng cao v\u00e0 3 \u0111i\u1ec1u c\u1ea7n ph\u1ea3i bi\u1ebft<\/strong>"},"content":{"rendered":"

PCR <\/a><\/strong>l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c ph\u00e1t tri\u1ec3n v\u00e0o nh\u1eefng n\u0103m 1970, m\u1ee5c ti\u00eau ch\u1ee7 y\u1ebfu l\u00e0 nh\u00e2n b\u1ea3n DNA m\u1ee5c ti\u00eau th\u00e0nh nhi\u1ec1u c\u00e1c b\u1ea3n sao. Trong v\u00e0i th\u1eadp k\u1ef7 qua, PCR truy\u1ec1n th\u1ed1ng \u0111\u00e3 \u0111\u01b0\u1ee3c c\u1ea3i ti\u1ebfn th\u00e0nh c\u00e1c lo\u1ea1i k\u1ef9 thu\u1eadt \u0111a d\u1ea1ng, hi\u1ec7n nay c\u00f3 h\u01a1n 30 bi\u1ebfn th\u1ec3 c\u1ee7a k\u1ef9 thu\u1eadt PCR (Rajalakshmi, 2017). Trong \u0111\u00f3, nested PCR l\u00e0 m\u1ed9t trong k\u1ef9 thu\u1eadt bi\u1ebfn th\u1ec3 c\u00f3 kh\u1ea3 n\u0103ng gia t\u0103ng t\u00ednh \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng.<\/p>\n\n\n\n

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V\u1eady Nested PCR l\u00e0 k\u1ef9 thu\u1eadt nh\u01b0 th\u1ebf n\u00e0o?<\/strong><\/h2>\n\n\n\n

Nested PCR l\u00e0m gi\u1ea3m s\u1ef1 khu\u1ebfch \u0111\u1ea1i s\u1ea3n ph\u1ea9m kh\u00f4ng \u0111\u1eb7c hi\u1ec7u b\u1eb1ng vi\u1ec7c th\u1ef1c hi\u1ec7n hai ph\u1ea3n \u1ee9ng PCR li\u00ean ti\u1ebfp. Ph\u1ea3n \u1ee9ng \u0111\u1ea7u ti\u00ean (PCR v\u00f2ng ngo\u00e0i) \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n v\u1edbi c\u1eb7p m\u1ed3i bao ph\u1ee7 \u0111o\u1ea1n tr\u00ecnh t\u1ef1 b\u1ed5 sung \u1edf hai \u0111\u1ea7u c\u1ee7a gen m\u1ee5c ti\u00eau. Nh\u01b0 v\u1eady, s\u1ea3n ph\u1ea9m c\u1ee7a ph\u1ea3n \u1ee9ng \u0111\u1ea7u ti\u00ean n\u00e0y l\u00e0 v\u00f9ng c\u00f3 ch\u1ee9a DNA m\u1ee5c ti\u00eau. Sau \u0111\u00f3 s\u1ea3n ph\u1ea9m c\u1ee7a ph\u1ea3n \u1ee9ng \u0111\u1ea7u ti\u00ean \u0111\u01b0\u1ee3c d\u00f9ng l\u00e0m tr\u00ecnh t\u1ef1 khu\u00f4n cho ph\u1ea3n \u1ee9ng PCR th\u1ee9 hai (PCR v\u00f2ng trong) , ph\u1ea3n \u1ee9ng n\u00e0y \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n v\u1edbi c\u1eb7p m\u1ed3i g\u1eafn \u0111\u1eb7c hi\u1ec7u v\u1edbi \u0111o\u1ea1n ch\u1ec9 ch\u1ee9a tr\u00ecnh t\u1ef1 m\u1ee5c ti\u00eau v\u00e0 t\u0103ng \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ea3u ph\u1ea3n \u1ee9ng.<\/p>\n\n\n\n

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\"K\u1ef9
H\u00ecnh 1. K\u1ef9 thu\u1eadt Nested PCR (Shen, 2019<\/a>)<\/em><\/figcaption><\/figure>\n\n\n\n

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C\u00e1c lo\u1ea1i nested PCR<\/strong><\/h2>\n\n\n\n

K\u1ef9 thu\u1eadt n\u00e0y chia l\u00e0m hai lo\u1ea1i c\u01a1 b\u1ea3n: double-tube v\u00e0 single-tube nested PCR. Double-tube nested PCR l\u00e0 k\u1ef9 thu\u1eadt th\u1ef1c hi\u1ec7n hai ph\u1ea3n \u1ee9ng PCR \u1edf hai tube kh\u00e1c nhau, s\u1ea3n ph\u1ea9m \u1edf tube PCR l\u1ea7n th\u1ee9 nh\u1ea5t s\u1ebd \u0111\u01b0\u1ee3c d\u00f9ng l\u00e0m m\u1eabu \u0111\u1ea7u v\u00e0o cho ph\u1ea3n \u1ee9ng PCR ti\u1ebfp theo \u1edf tube th\u1ee9 hai. Lo\u1ea1i n\u00e0y gi\u00fap l\u00e0m gia t\u0103ng \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u0169ng nh\u01b0 \u0111\u1ed9 nh\u1ea1y ph\u1ea3n \u1ee9ng, \u0111\u1eb7c bi\u1ec7t l\u00e0 \u0111\u1ed1i v\u1edbi c\u1eb7p m\u1ed3i c\u00f3 \u0111\u1ed9 nh\u1ea1y th\u1ea5p. Tuy nhi\u00ean, nguy c\u01a1 ngo\u1ea1i nhi\u1ec5m l\u1ea1i r\u1ea5t cao khi s\u1eed d\u1ee5ng lo\u1ea1i double-tube, do ph\u1ea3i m\u1edf n\u1eafp \u1edf c\u1ea3 hai tube khi th\u00eam s\u1ea3n ph\u1ea9m PCR v\u00f2ng ngo\u00e0i v\u00e0o tube PCR v\u00f2ng trong.<\/p>\n\n\n\n

\u0110\u1ed1i v\u1edbi lo\u1ea1i single-tube, c\u1ea3 hai ph\u1ea3n \u1ee9ng PCR \u0111\u01b0\u1ee3c k\u1ebft h\u1ee3p v\u00e0o c\u00f9ng m\u1ed9t tube (hay c\u00f2n g\u1ecdi l\u00e0 non-stop nested PCR), ngh\u0129a l\u00e0 sau khi ph\u1ea3n \u1ee9ng PCR v\u00f2ng ngo\u00e0i di\u1ec5n ra th\u00ec kh\u00f4ng c\u1ea7n d\u1eebng l\u1ea1i l\u1ea5y s\u1ea3n ph\u1ea9m m\u00e0 ti\u1ebfp t\u1ee5c th\u1ef1c hi\u1ec7n PCR v\u00f2ng trong ngay sau \u0111\u00f3 m\u00e0 kh\u00f4ng c\u1ea7n \u0111\u1ed5i tube. Lo\u1ea1i single-tube gi\u00fap h\u1ea1n ch\u1ebf nguy c\u01a1 nhi\u1ec5m ch\u00e9o c\u0169ng nh\u01b0 gi\u1ea3m th\u1eddi gian th\u1ef1c hi\u1ec7n ph\u1ea3n \u1ee9ng h\u01a1n so v\u1edbi double-tube. <\/p>\n\n\n\n

\u0110\u1ec3 th\u1ef1c hi\u1ec7n \u0111\u01b0\u1ee3c single-tube th\u00ec m\u1ed3i cho ph\u1ea3n \u1ee9ng PCR v\u00f2ng ngo\u00e0i (outer primers) \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf sao cho c\u00f3 nhi\u1ec7t \u0111\u1ed9 b\u1eaft c\u1eb7p cao h\u01a1n m\u1ed3i c\u1ee7a ph\u1ea3n \u1ee9ng PCR v\u00f2ng trong (inner primers) t\u1eeb 8-10o<\/sup>C, \u0111i\u1ec1u n\u00e0y gi\u00fap gi\u1ea3m s\u1ef1 b\u1eaft c\u1eb7p kh\u00f4ng \u0111\u1eb7c hi\u1ec7u c\u1ee7a c\u00e1c c\u1eb7p m\u1ed3i \u1edf m\u1ed7i ph\u1ea3n \u1ee9ng PCR. N\u1ed3ng \u0111\u1ed9 c\u1eb7p m\u1ed3i v\u00f2ng ngo\u00e0i c\u1ea7n \u0111\u01b0\u1ee3c t\u1ed1i \u01b0u tr\u00e1nh d\u01b0 th\u1eeba. Ngo\u00e0i ra, nested PCR c\u00f2n m\u1ed9t bi\u1ebfn th\u1ec3 kh\u00e1c l\u00e0 semi-nested PCR, lo\u1ea1i n\u00e0y s\u1eed d\u1ee5ng m\u1ed9t m\u1ed3i v\u00f2ng ngo\u00e0i l\u00e0m m\u1ed9t trong hai m\u1ed3i c\u1ee7a c\u1eb7p m\u1ed3i v\u00f2ng trong hay n\u00f3i c\u00e1ch kh\u00e1c l\u00e0 k\u1ef9 thu\u1eadt n\u00e0y ch\u1ec9 s\u1eed d\u1ee5ng ba m\u1ed3i cho c\u1ea3 hai ph\u1ea3n \u1ee9ng.<\/p>\n\n\n\n

\"K\u1ef9
H\u00ecnh 2. K\u1ef9 thu\u1eadt Semi-nested PCR (Shen, 2019)<\/em><\/figcaption><\/figure>\n\n\n\n

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\u1ee8ng d\u1ee5ng c\u1ee7a nested PCR<\/strong><\/h2>\n\n\n\n

Nh\u01b0 v\u1eady, nested PCR l\u00e0 k\u1ef9 thu\u1eadt l\u00e0m t\u0103ng \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u v\u00e0 \u0111\u1ed9 nh\u1ea1y khi nh\u00e2n b\u1ea3n s\u1ea3n ph\u1ea9m DNA m\u1ee5c ti\u00eau b\u1eb1ng c\u00e1ch th\u1ef1c hi\u1ec7n hai ph\u1ea3n \u1ee9ng t\u1ed5ng h\u1ee3p. Th\u00eam v\u00e0o \u0111\u00f3 \u0111\u1ec3 gia t\u0103ng hi\u1ec7u qu\u1ea3 nh\u00e2n b\u1ea3n th\u00ec nested PCR c\u0169ng \u0111\u01b0\u1ee3c k\u1ebft h\u1ee3p v\u1edbi nhi\u1ec1u k\u1ef9 thu\u1eadt PCR kh\u00e1c nh\u01b0 real-time PCR hay multiplex PCR. Hi\u1ec7n nay, k\u1ef9 thu\u1eadt n\u00e0y \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng ph\u1ed5 bi\u1ebfn trong ph\u00e2n t\u00edch ph\u00e1t sinh gene (phylogenetic analysis) hay \u0111a h\u00ecnh di truy\u1ec1n (genetic polymorphism) \u1ee9ng d\u1ee5ng trong vi\u1ec7c ph\u00e1t hi\u1ec7n c\u00e1c gene g\u00e2y b\u1ec7nh hay \u0111\u1ed9t bi\u1ebfn.<\/p>\n\n\n\n

M\u1ed9t \u1ee9ng d\u1ee5ng hi\u1ec7u qu\u1ea3 c\u1ee7a nested PCR \u0111\u00f3 l\u00e0 x\u00e1c \u0111\u1ecbnh b\u1ec7nh \u0111\u1ed1m tr\u1eafng tr\u00ean t\u00f4m trong th\u1eddi gian ng\u1eafn. \u1ede Vi\u1ec7t Nam c\u0169ng nh\u01b0 tr\u00ean th\u1ebf gi\u1edbi th\u00ec vi\u1ec7c nu\u00f4i tr\u1ed3ng t\u00f4m \u0111ang c\u00f3 xu h\u01b0\u1edbng ph\u00e1t tri\u1ec3n nhanh. Tuy nhi\u00ean \u0111i \u0111\u00f4i v\u1edbi s\u1ef1 ph\u00e1t tri\u1ec3n \u0111\u00f3 l\u00e0 s\u1ef1 b\u00f9ng ph\u00e1t c\u1ee7a c\u00e1c d\u1ecbch b\u1ec7nh g\u00e2y h\u1ea1i cho t\u00f4m \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn ch\u1ea5t l\u01b0\u1ee3ng c\u0169ng nh\u01b0 t\u1ed5n th\u1ea5t n\u1eb7ng n\u1ec1 cho ng\u01b0\u1eddi nu\u00f4i tr\u1ed3ng. <\/p>\n\n\n\n

\u0110\u1eb7c bi\u1ec7t t\u00f4m r\u1ea5t hay b\u1ecb t\u1ea5n c\u00f4ng b\u1edfi c\u00e1c lo\u1ea1i virus nh\u01b0 virus g\u00e2y b\u1ec7nh \u0111\u1ed1m tr\u1eafng (white spot syndrome virus \u2013 WSSV), virus IHHNV (Infectious Hypothermal and Hematopoietic Necrosis Virus, IHHNV), virus g\u00e2y h\u1ed9i ch\u1ee9ng Taura (Taura Syndrome Virus \u2013 TSV), b\u1ec7nh ho\u1ea1i t\u1eed tuy\u1ebfn ru\u1ed9t gi\u1eefa (Baculovirus Migut and Necrosis \u2013 BMN), \u2026 Trong \u0111\u00f3, lo\u00e0i WSSV \u0111\u01b0\u1ee3c coi l\u00e0 \u201cs\u00e1t th\u1ee7\u201d nguy hi\u1ec3m \u1edf c\u00e1c ao nu\u00f4i t\u00f4m, do n\u00f3 c\u00f3 th\u1ec3 g\u00e2y t\u1ef7 l\u1ec7 ch\u1ebft khi b\u1ecb nhi\u1ec5m l\u00ean \u0111\u1ebfn 80 \u2013 100% sau kho\u1ea3ng 10 ng\u00e0y nhi\u1ec5m. <\/p>\n\n\n\n

Do v\u1eady nested PCR gi\u00fap ch\u1ea9n \u0111o\u00e1n nhanh ch\u00f3ng m\u1ea7m b\u1ec7nh \u1edf ao nu\u00f4i t\u00f4m, t\u1eeb \u0111\u00f3 gi\u1ea3m thi\u1ec7t h\u1ea1i cho ng\u01b0\u1eddi nu\u00f4i. C\u00e1c nghi\u00ean c\u1ee9u \u1edf \u0110\u1ea1i h\u1ecdc C\u1ea7n Th\u01a1 th\u1ef1c \u0111\u00e3 s\u1eed d\u1ee5ng nested PCR \u0111\u1ec3 ph\u00e1t hi\u1ec7n WSSV tr\u00ean t\u00f4m v\u00e0 c\u00f3 nh\u1eefng \u0111\u00e1nh gi\u00e1 v\u1ec1 quy tr\u00ecnh (T.T. Tuy\u1ebft Hoa, 2011). Quy tr\u00ecnh n\u00e0y \u0111\u00e3 cho th\u1ea5y hi\u1ec7u qu\u1ea3 trong vi\u1ec7c ph\u00e1t hi\u1ec7n WSSV t\u1eeb nhi\u1ec1u \u0111\u1ed1i t\u01b0\u1ee3ng c\u1ea3m nhi\u1ec5m, v\u1edbi \u0111\u1ed9 nh\u1ea1y cao v\u00e0 th\u1eddi gian khu\u1ebfch \u0111\u1ea1i kho\u1ea3ng hai gi\u1edd. M\u1ed9t nghi\u00ean c\u1ee9u t\u01b0\u01a1ng t\u1ef1 c\u0169ng \u0111\u01b0\u1ee3c th\u1ef1c v\u1edbi m\u1eabu t\u00f4m t\u1eeb C\u00e0 Mau, nested PCR \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng v\u00e0 mang l\u1ea1i \u0111\u1ed9 ch\u00ednh x\u00e1c v\u00e0 \u0111\u1ed9 nh\u1ea1y cao trong th\u1eddi gian ng\u1eafn.<\/p>\n\n\n\n

\"Virus
H\u00ecnh 3. Virus \u0111\u1ed1m tr\u1eafng (White Spot Syndrome Virus – WSSV) g\u00e2y b\u1ec7nh tr\u00ean t\u00f4m<\/em><\/figcaption><\/figure>\n\n\n\n

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Ngo\u00e0i b\u1ec7nh tr\u00ean t\u00f4m, nested PCR c\u0169ng \u0111\u01b0\u1ee3c nghi\u00ean c\u1ee9u \u1ee9ng d\u1ee5ng trong vi\u1ec7c chu\u1ea9n \u0111o\u00e1n ph\u00e1t hi\u1ec7n v\u00e0 x\u00e1c \u0111\u1ecbnh type porcine circovirus type 2 (PCV2) g\u00e2y ra nhi\u1ec1u h\u1ed9i ch\u1ee9ng b\u1ec7nh \u1edf l\u1ee3n nh\u01b0 h\u1ed9i ch\u1ee9ng g\u1ea7y c\u00f2m \u1edf l\u1ee3n sau cai s\u1eefa (Postweaning Multisystemic Wasting Syndrome \u2013 PMWS), h\u1ed9i ch\u1ee9ng vi\u00eam da v\u00e0 vi\u00eam th\u1eadn (Porcine Dermatitis and Nephropathy Syndrome \u2013 PDNS), h\u1ed9i ch\u1ee9ng vi\u00eam \u0111\u01b0\u1eddng h\u00f4 h\u1ea5p (Porcine Respiratory Diseases Complex) v\u00e0 h\u1ed9i ch\u1ee9ng r\u1ed1i lo\u1ea1n sinh s\u1ea3n (Porcine Reproductive Disorders) (Opriessnig, 2007). Nghi\u00ean c\u1ee9u cho th\u1ea5y ph\u1ea3n \u1ee9ng nested PCR gi\u00fap x\u00e1c \u0111\u1ecbnh s\u1ef1 c\u00f3 m\u1eb7t c\u1ee7a PCV2 t\u01b0\u01a1ng \u1ee9ng v\u1edbi tri\u1ec7u ch\u1ee9ng l\u00e2m s\u00e0ng c\u1ee7a l\u1ee3n (c\u00f3\/kh\u00f4ng c\u00f3 tri\u1ec7u ch\u1ee9ng b\u1ec7nh) (HT M\u1ef9 L\u1ec7, 2012).<\/p>\n\n\n\n

\u0110\u1ed1i v\u1edbi ch\u1ea9n \u0111o\u00e1n b\u1ec7nh tr\u00ean ng\u01b0\u1eddi, Roy v\u00e0 c\u1ed9ng s\u1ef1 (2021) \u0111\u00e3 th\u1ef1c hi\u1ec7n \u0111\u00e1nh gi\u00e1 hi\u1ec7u qu\u1ea3 ch\u1ea9n \u0111o\u00e1n b\u1ec7nh s\u1ed1t ph\u00e1t ban b\u1eb1ng nested PCR v\u00e0 k\u1ef9 thu\u1eadt LAMP. K\u1ebft qu\u1ea3 \u0111\u00e3 ph\u00e1t hi\u1ec7n DNA c\u1ee7a vi khu\u1ea9n Orientia tsutsugamushi<\/em> b\u1eb1ng c\u00e1ch nh\u1eafm m\u1ee5c ti\u00eau gen 47-kDa. K\u1ebft qu\u1ea3 nghi\u00ean c\u1ee9u ghi nh\u1eadn \u0111\u01b0\u1ee3c v\u1edbi k\u1ef9 thu\u1eadt nested PCR cho \u0111\u1ed9 nh\u1ea1y 29,73% trong khi LAMP l\u00e0 16,22%. \u0110\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a nested PCR v\u00e0 LAMP r\u1ea5t cao, l\u1ea7n l\u01b0\u1ee3t l\u00e0 99,58% v\u00e0 99,16%. \u0110\u1ed9 ch\u00ednh x\u00e1c ch\u1ea9n \u0111o\u00e1n c\u1ee7a PCR l\u1ed3ng nhau (90,15%) \u0111\u01b0\u1ee3c cho l\u00e0 t\u1ed1t h\u01a1n \u0111\u00f4i ch\u00fat so v\u1edbi LAMP (87,96%).<\/p>\n\n\n\n

K\u1ef9 thu\u1eadt nested PCR k\u1ebft h\u1ee3p multiplex PCR c\u0169ng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 gia t\u0103ng hi\u1ec7u qu\u1ea3 trong x\u00e1c \u0111\u1ecbnh ki\u1ec3u gene c\u1ee7a HPV (Mudhiget, 2022). C\u00e1c plasmid ch\u1ee9a HPV-DNA v\u00e0 m\u1eabu m\u00f4 ung th\u01b0 c\u1ed5 t\u1eed cung \u0111\u01b0\u1ee3c d\u00f9ng \u0111\u1ec3 \u0111\u00e1nh gi\u00e1 nested-multiplex PCR (NM-PCR). Trong PCR v\u00f2ng \u0111\u1ea7u, c\u00e1c c\u1eb7p m\u1ed3i \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf \u0111\u1ec3 khu\u1ebfch \u0111\u1ea1i 38 lo\u1ea1i HPV. \u1ede PCR v\u00f2ng hai, c\u00e1c \u0111o\u1ea1n m\u1ed3i \u0111\u1eb7c hi\u1ec7u \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng cho 15 lo\u1ea1i HPV. K\u1ebft qu\u1ea3 ghi nh\u1eadn \u0111\u01b0\u1ee3c cho th\u1ea5y NM-PCR c\u00f3 th\u1ec3 khu\u1ebfch \u0111\u1ea1i t\u1ea5t c\u1ea3 c\u00e1c lo\u1ea1i HPV t\u1eeb 75 m\u1eabu, k\u1ef9 thu\u1eadt n\u00e0y h\u01b0\u1edbng \u0111\u1ebfn vi\u1ec7c tr\u1edf th\u00e0nh ph\u01b0\u01a1ng ph\u00e1p h\u1ee9a h\u1eb9n \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh ch\u00ednh x\u00e1c ki\u1ec3u gen c\u1ee7a HPV.<\/p>\n\n\n\n

M\u1ed9t c\u00f4ng b\u1ed1 kh\u00e1c c\u0169ng ghi nh\u1eadn hi\u1ec7u qu\u1ea3 c\u1ee7a vi\u1ec7c s\u1eed d\u1ee5ng k\u1ef9 thu\u1eadt semi-nested PCR \u0111\u1ec3 ph\u00e1t hi\u1ec7n c\u00e1c ch\u1ee7ng Helicobactor pylori<\/em> mang gene CagA tr\u1ef1c ti\u1ebfp t\u1eeb m\u1eabu d\u1ecbch d\u1ea1 d\u00e0y (Nguy\u1ec5n Ph\u00fa H\u00f9ng, 2020) . Nghi\u00ean c\u1ee9u h\u01b0\u1edbng \u0111\u1ebfn t\u1ea7m so\u00e1t ung th\u01b0 d\u1ea1 d\u00e0y c\u0169ng nh\u01b0 gi\u1ea3m t\u00ednh x\u00e2m l\u1ea5n so v\u1edbi c\u00e1c ph\u01b0\u01a1ng ph\u00e1p ph\u00e2n t\u00edch t\u1eeb m\u00f4 d\u1ea1 d\u00e0y. Ph\u00e2n t\u00edch tr\u00ean 35 m\u1eabu b\u1ec7nh ph\u1ea9m b\u1eb1ng nested PCR \u0111\u00e3 cho th\u1ea5y t\u1ef7 l\u1ec7 c\u00e1c m\u1eabu d\u01b0\u01a1ng t\u00ednh v\u1edbi ch\u1ee7ng mang gene CagA chi\u1ebfm t\u1ef7 l\u1ec7 cao, l\u00ean t\u1edbi 90%.<\/p>\n\n\n\n

K\u1ef9 thu\u1eadt nested real-time PCR c\u0169ng \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng \u0111\u1ec3 nghi\u00ean c\u1ee9u nh\u1eb1m x\u00e1c \u0111\u1ecbnh gene m\u00e3 h\u00f3a ch\u1ea5t g\u00e2y d\u1ecb \u1ee9ng c\u00f3 trong c\u00e1c th\u1ef1c ph\u1ea9m c\u00f3 h\u1ea1t \u0111i\u1ec1u (Costa v\u00e0 c\u1ed9ng s\u1ef1, 2022). K\u1ef9 thu\u1eadt n\u00e0y \u0111\u1ea1t \u0111\u1ed9 nh\u1ea1y cao v\u1edbi 10 mg\/kg h\u1ea1t \u0111i\u1ec1u trong b\u1ed9t\/b\u00e1nh quy m\u00e0 kh\u00f4ng b\u1ecb \u1ea3nh h\u01b0\u1edfng b\u1edfi nhi\u1ec7t khi x\u1eed l\u00fd b\u00e1nh. V\u00ec v\u1eady nested real-time PCR \u0111\u01b0\u1ee3c \u0111\u1ec1 xu\u1ea5t l\u00e0m c\u00f4ng c\u1ee5 \u0111\u1ecbnh l\u01b0\u1ee3ng\/x\u00e1c nh\u1eadn \u0111\u1ec3 ph\u00e2n t\u00edch ch\u1ea5t g\u00e2y d\u1ecb \u1ee9ng trong th\u1ef1c ph\u1ea9m ch\u1ee9a h\u1ea1t \u0111i\u1ec1u.<\/p>","protected":false},"excerpt":{"rendered":"

PCR l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c ph\u00e1t tri\u1ec3n v\u00e0o nh\u1eefng n\u0103m 1970, m\u1ee5c ti\u00eau ch\u1ee7 y\u1ebfu l\u00e0 nh\u00e2n b\u1ea3n DNA m\u1ee5c ti\u00eau th\u00e0nh nhi\u1ec1u c\u00e1c b\u1ea3n sao. Trong v\u00e0i th\u1eadp k\u1ef7 qua, PCR truy\u1ec1n th\u1ed1ng \u0111\u00e3 \u0111\u01b0\u1ee3c c\u1ea3i ti\u1ebfn th\u00e0nh c\u00e1c lo\u1ea1i k\u1ef9 thu\u1eadt \u0111a d\u1ea1ng, hi\u1ec7n nay c\u00f3 h\u01a1n 30 bi\u1ebfn th\u1ec3 c\u1ee7a k\u1ef9 thu\u1eadt PCR (Rajalakshmi, 2017). Trong \u0111\u00f3, nested PCR l\u00e0 m\u1ed9t trong k\u1ef9 thu\u1eadt bi\u1ebfn th\u1ec3 c\u00f3 kh\u1ea3 n\u0103ng gia t\u0103ng t\u00ednh \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng.<\/p>","protected":false},"author":1,"featured_media":7484,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_uag_custom_page_level_css":"","site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"categories":[267,228],"tags":[286],"class_list":["post-7479","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-sinh-hoc-phan-tu-bai-viet","category-bai-viet","tag-nested-pcr"],"uagb_featured_image_src":{"full":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-scaled.jpg",1920,1078,false],"thumbnail":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-150x150.jpg",150,150,true],"medium":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-300x168.jpg",300,168,true],"medium_large":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-768x431.jpg",768,431,true],"large":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-1024x575.jpg",1024,575,true],"1536x1536":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-1536x862.jpg",1536,862,true],"2048x2048":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-2048x1149.jpg",2048,1149,true],"trp-custom-language-flag":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-18x10.jpg",18,10,true],"woocommerce_thumbnail":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-scaled-1707x958.jpg",1707,958,true],"woocommerce_single":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-2560x1437.jpg",2560,1437,true],"woocommerce_gallery_thumbnail":["https:\/\/abtvn.com\/wp-content\/uploads\/2023\/01\/DSCF5072-scaled-100x100.jpg",100,100,true]},"uagb_author_info":{"display_name":"ABT Team","author_link":"https:\/\/abtvn.com\/en\/author\/admin\/"},"uagb_comment_info":0,"uagb_excerpt":"PCR l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c ph\u00e1t tri\u1ec3n v\u00e0o nh\u1eefng n\u0103m 1970, m\u1ee5c ti\u00eau ch\u1ee7 y\u1ebfu l\u00e0 nh\u00e2n b\u1ea3n DNA m\u1ee5c ti\u00eau th\u00e0nh nhi\u1ec1u c\u00e1c b\u1ea3n sao. Trong v\u00e0i th\u1eadp k\u1ef7 qua, PCR truy\u1ec1n th\u1ed1ng \u0111\u00e3 \u0111\u01b0\u1ee3c c\u1ea3i ti\u1ebfn th\u00e0nh c\u00e1c lo\u1ea1i k\u1ef9 thu\u1eadt \u0111a d\u1ea1ng, hi\u1ec7n nay c\u00f3 h\u01a1n 30 bi\u1ebfn th\u1ec3 c\u1ee7a k\u1ef9…","_links":{"self":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/7479","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/comments?post=7479"}],"version-history":[{"count":4,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/7479\/revisions"}],"predecessor-version":[{"id":7855,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/7479\/revisions\/7855"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/media\/7484"}],"wp:attachment":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/media?parent=7479"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/categories?post=7479"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/tags?post=7479"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}