{"id":9888,"date":"2024-03-05T10:54:18","date_gmt":"2024-03-05T03:24:18","guid":{"rendered":"https:\/\/abtvn.com\/?p=9888"},"modified":"2025-10-22T16:13:21","modified_gmt":"2025-10-22T08:43:21","slug":"su-khac-nhau-giua-pcr-va-real-time-pcr","status":"publish","type":"post","link":"https:\/\/abtvn.com\/en\/su-khac-nhau-giua-pcr-va-real-time-pcr\/","title":{"rendered":"S\u1ef1 kh\u00e1c nhau gi\u1eefa PCR v\u00e0 Real-time PCR<\/strong>"},"content":{"rendered":"
\n\t\t\t\t
\n\t\t\t\t\t
\n\t\t\t\t
\n\t\t\t\t
\n\t\t\t\t\t\t\t\t\t\n

1. Th\u1ebf n\u00e0o l\u00e0 PCR?<\/strong><\/h2>\n\n

PCR (Polymerase Chain Reaction)<\/a> <\/strong>l\u00e0 k\u1ef9 thu\u1eadt sinh h\u1ecdc ph\u00e2n t\u1eed nh\u00e2n b\u1ea3n gene m\u1ee5c ti\u00eau l\u00ean h\u00e0ng tri\u1ec7u l\u1ea7n trong m\u00f4i tr\u01b0\u1eddng in vitro <\/em>d\u1ef1a v\u00e0o s\u1ef1 thay \u0111\u1ed5i nhi\u1ec7t \u0111\u1ed9 theo chu k\u1ef3, qu\u00e1 tr\u00ecnh nh\u00e2n b\u1ea3n \u0111\u01b0\u1ee3c m\u00f4 ph\u1ecfng theo c\u01a1 ch\u1ebf c\u1ee7a qu\u00e1 tr\u00ecnh sao ch\u00e9p DNA trong t\u1ebf b\u00e0o (Putra v\u00e0 c\u1ed9ng s\u1ef1, 2020). K\u1ef9 thu\u1eadt n\u00e0y c\u00f3 kh\u1ea3 n\u0103ng ph\u00e1t hi\u1ec7n ch\u00ednh x\u00e1c m\u1ee5c ti\u00eau v\u1edbi l\u01b0\u1ee3ng m\u1eabu t\u1ed1i thi\u1ec3u, m\u00e0 c\u00e1c ph\u01b0\u01a1ng ph\u00e1p kh\u00e1c nh\u01b0 nu\u00f4i c\u1ea5y ho\u1eb7c huy\u1ebft thanh h\u1ecdc kh\u00f4ng th\u1ec3 th\u1ef1c hi\u1ec7n \u0111\u01b0\u1ee3c. \u0110\u1ed9 \u0111\u1eb7c hi\u1ec7u v\u00e0 \u0111\u1ed9 nh\u1ea1y cao c\u1ee7a PCR l\u00e0 l\u00fd do khi\u1ebfn n\u00f3 \u0111\u01b0\u1ee3c coi l\u00e0 \u201cti\u00eau chu\u1ea9n v\u00e0ng\u201d trong c\u00e1c x\u00e9t nghi\u1ec7m ch\u1ea9n \u0111o\u00e1n b\u1ec7nh.<\/p>\n\n

\u00a0<\/div>\n\n

Nguy\u00ean t\u1eafc c\u1ee7a k\u1ef9 thu\u1eadt PCR<\/em><\/strong><\/h3>\n\n

C\u01a1 s\u1edf c\u1ee7a PCR l\u00e0 d\u1ef1a tr\u00ean qu\u00e1 tr\u00ecnh sao ch\u00e9p DNA trong t\u1ebf b\u00e0o, c\u1ea3 hai \u0111\u1ec1u l\u00e0 t\u1ed5ng h\u1ee3p DNA nh\u01b0ng v\u1ec1 \u0111\u1eb7c t\u00ednh c\u00f3 s\u1ef1 kh\u00e1c bi\u1ec7t r\u00f5 r\u00e0ng. Sao ch\u00e9p DNA l\u00e0 qu\u00e1 tr\u00ecnh sinh h\u1ecdc t\u1ea1o ra hai b\u1ea3n sao DNA gi\u1ed1ng h\u1ec7t nhau t\u1eeb m\u1ed9t ph\u00e2n t\u1eed DNA ban \u0111\u1ea7u hay c\u00f2n g\u1ecdi l\u00e0 t\u1ed5ng h\u1ee3p gene in vivo. <\/em>Qu\u00e1 tr\u00ecnh n\u00e0y mang t\u00ednh li\u00ean t\u1ee5c v\u00e0 c\u1ea7n \u0111\u1ed9 ch\u00ednh x\u00e1c cao, m\u1ee5c \u0111\u00edch c\u1ee7a sao ch\u00e9p \u0111\u1ec3 nh\u00e2n b\u1ea3n to\u00e0n h\u1ec7 gene cho s\u1ef1 ph\u00e2n chia t\u1ebf b\u00e0o. Trong khi \u0111\u00f3, PCR l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt sinh h\u1ecdc ph\u00e2n t\u1eed \u0111\u1ec3 t\u1ea1o ra nhi\u1ec1u b\u1ea3n sao c\u1ee7a m\u1ed9t \u0111o\u1ea1n DNA c\u1ee5 th\u1ec3 \u1edf \u0111i\u1ec1u ki\u1ec7n in vitro, <\/em>k\u1ef9 thu\u1eadt n\u00e0y di\u1ec5n ra trong th\u1eddi gian nh\u1ea5t \u0111\u1ecbnh (theo s\u1ed1 chu k\u1ef3) v\u00e0 d\u1ec5 b\u1ecb \u1ea3nh h\u01b0\u1edfng t\u1eeb c\u00e1c th\u00e0nh ph\u1ea7n ph\u1ea3n \u1ee9ng c\u0169ng nh\u01b0 \u0111i\u1ec1u ki\u1ec7n b\u00ean ngo\u00e0i.<\/p>\n\n

Th\u00e0nh ph\u1ea7n c\u01a1 b\u1ea3n m\u1ed9t ph\u1ea3n \u1ee9ng PCR bao g\u1ed3m: DNA khu\u00f4n, c\u1eb7p m\u1ed3i, DNA polymerase, dNTP v\u00e0 dung d\u1ecbch \u0111\u1ec7m. \u1ed0ng ph\u1ea3n \u1ee9ng sau khi \u0111\u00e3 \u0111\u1ea3m b\u1ea3o \u0111\u1ee7 c\u00e1c th\u00e0nh ph\u1ea7n s\u1ebd \u0111\u1eb7t v\u00e0o m\u00e1y lu\u00e2n nhi\u1ec7t (hay c\u00f2n g\u1ecdi l\u00e0 m\u00e1y PCR) \u0111\u00e3 c\u00e0i s\u1eb5n chu tr\u00ecnh nhi\u1ec7t. Qu\u00e1 tr\u00ecnh khu\u1ebfch \u0111\u1ea1i s\u1ebd tr\u1ea3i qua c\u00e1c giai \u0111o\u1ea1n gia gi\u1ea3m nhi\u1ec7t theo chu k\u1ef3 \u0111\u1ec3 ti\u1ebfn h\u00e0nh khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n gene m\u1ee5c ti\u00eau.<\/p>\n\n

\"H\u00ecnh\n
Figure 1. <\/strong>Quy tr\u00ecnh PCR<\/em> (Polymerase Chain Reaction)<\/em><\/figcaption>\n<\/figure>\n\n

2. Th\u1ebf n\u00e0o l\u00e0 Real-time PCR?<\/strong><\/h2>\n\n

V\u1eady th\u1ebf n\u00e0o l\u00e0 Real-time PCR? Real-time PCR l\u00e0 k\u1ef9 thu\u1eadt c\u1ea3i ti\u1ebfn t\u1eeb PCR, cho ph\u00e9p theo d\u00f5i s\u1ef1 gia t\u0103ng v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng DNA m\u1ee5c ti\u00eau trong m\u1eabu th\u1eed trong su\u1ed1t th\u1eddi gian th\u1ef1c t\u1ebf di\u1ec5n ra ph\u1ea3n \u1ee9ng nh\u1edd v\u00e0o c\u00f4ng ngh\u1ec7 ph\u00e1t hu\u1ef3nh quang (Higuchi v\u00e0 c\u1ed9ng s\u1ef1, 1993).<\/p>\n\n

\u00a0<\/div>\n\n

Nguy\u00ean t\u1eafc c\u1ee7a k\u1ef9 thu\u1eadt Real-time PCR<\/strong><\/h3>\n\n

K\u1ef9 thu\u1eadt Real-time PCR (hay Quantitative Polymerase Chain Reaction – qPCR) v\u1ec1 c\u01a1 b\u1ea3n v\u1eabn l\u00e0 PCR, do v\u1eady nguy\u00ean t\u1eafc v\u1eabn l\u00e0 d\u00f9ng c\u1eb7p m\u1ed3i \u0111\u1eb7c hi\u1ec7u \u0111\u1ec3 ph\u00e1t hi\u1ec7n v\u00e0 khu\u1ebfch \u0111\u1ea1i tr\u00ecnh t\u1ef1 m\u1ee5c ti\u00eau b\u1eb1ng c\u00e1ch thay \u0111\u1ed5i nhi\u1ec7t \u0111\u1ed9 theo chu k\u1ef3.<\/p>\n\n

\u0110i\u1ec3m kh\u00e1c bi\u1ec7t \u0111\u00f3 l\u00e0 Real-time PCR d\u1ef1a tr\u00ean vi\u1ec7c s\u1eed d\u1ee5ng c\u00e1c m\u1eabu d\u00f2 hay thu\u1ed1c nhu\u1ed9m ph\u00e1t hu\u1ef3nh quang g\u1eafn v\u1edbi DNA m\u1ee5c ti\u00eau \u0111\u1ec3 ch\u1ec9 th\u1ecb s\u1ef1 gia t\u0103ng b\u1ea3n sao DNA m\u1ee5c ti\u00eau, l\u01b0\u1ee3ng t\u00edn hi\u1ec7u ghi nh\u1eadn \u0111\u01b0\u1ee3c \u0111o trong th\u1eddi gian th\u1ef1c hi\u1ec7n ph\u1ea3n \u1ee9ng (Mackay, 2004).<\/em> Theo \u0111\u00f3, khi ph\u1ea3n \u1ee9ng nh\u00e2n b\u1ea3n x\u1ea3y ra t\u1edbi m\u1ed9t chu k\u1ef3 nh\u1ea5t \u0111\u1ecbnh, c\u01b0\u1eddng \u0111\u1ed9 t\u00edn hi\u1ec7u hu\u1ef3nh quang s\u1ebd b\u1eaft \u0111\u1ea7u c\u00f3 s\u1ef1 gia t\u0103ng r\u00f5 r\u1ec7t v\u00e0 t\u01b0\u01a1ng quan v\u1edbi s\u1ed1 l\u01b0\u1ee3ng b\u1ea3n sao DNA \u0111\u01b0\u1ee3c t\u1ea1o ra. K\u1ebft qu\u1ea3 \u0111\u01b0\u1ee3c th\u1ec3 hi\u1ec7n tr\u00ean m\u00e1y t\u00ednh d\u01b0\u1edbi d\u1ea1ng bi\u1ec3u \u0111\u1ed3 quan s\u00e1t \u0111\u01b0\u1ee3c qua m\u1ed7i chu k\u1ef3, t\u1eeb \u0111\u00f3 c\u00f3 th\u1ec3 \u0111\u01b0a ra \u0111\u00e1nh gi\u00e1 v\u1ec1 hi\u1ec7u qu\u1ea3 khu\u1ebfch \u0111\u1ea1i DNA m\u1ee5c ti\u00eau (H\u00ecnh 2)<\/em><\/strong>.<\/p>\n\n

Ngo\u00e0i ra, Real-time PCR y\u00eau c\u1ea7u c\u00f3 thi\u1ebft b\u1ecb \u0111o c\u01b0\u1eddng \u0111\u1ed9 ph\u00e1t hu\u1ef3nh quang t\u1eeb \u1ed1ng m\u1eabu v\u00e0 c\u00e0i ch\u01b0\u01a1ng tr\u00ecnh ph\u1ea7n m\u1ec1m cho ph\u00e9p x\u1eed l\u00fd k\u1ebft qu\u1ea3 v\u1ec1 s\u1ef1 bi\u1ebfn \u0111\u1ed5i c\u01b0\u1eddng \u0111\u1ed9 hu\u1ef3nh quang.<\/p>\n\n

\"H\u00ecnh\n
Figure 2. <\/strong>Quy tr\u00ecnh Real-time PCR<\/em><\/figcaption>\n<\/figure>\n\n

3. S\u1ef1 t\u01b0\u01a1ng \u0111\u1ed3ng gi\u1eefa PCR v\u00e0 qPCR<\/strong><\/h2>\n\n

V\u1ec1 b\u1ea3n ch\u1ea5t, qPCR ch\u00ednh l\u00e0 PCR \u0111\u01b0\u1ee3c c\u1ea3i ti\u1ebfn, do v\u1eady s\u1ebd c\u00f3 nh\u1eefng \u0111i\u1ec3m t\u01b0\u01a1ng \u0111\u1ed3ng gi\u1eefa hai k\u1ef9 thu\u1eadt n\u00e0y. Theo \u0111\u00f3, c\u1ea3 hai k\u1ef9 thu\u1eadt n\u00e0y \u0111\u1ec1u d\u00f9ng \u0111\u1ec3 nh\u00e2n b\u1ea3n tr\u00ecnh t\u1ef1 gene c\u1ee5 th\u1ec3 \u0111\u01b0\u1ee3c gi\u1edbi h\u1ea1n b\u1edfi c\u1eb7p m\u1ed3i \u0111\u1eb7c hi\u1ec7u, gi\u00fap ph\u00e1t hi\u1ec7n \u0111\u01b0\u1ee3c t\u00e1c nh\u00e2n m\u1ee5c ti\u00eau trong x\u00e9t nghi\u1ec7m (\u0111\u1ecbnh t\u00ednh). M\u1ed9t \u0111i\u1ec3m t\u01b0\u01a1ng \u0111\u1ed3ng n\u1eefa c\u1ee7a hai k\u1ef9 thu\u1eadt n\u00e0y l\u00e0 \u0111\u1ec1u s\u1eed d\u1ee5ng DNA l\u00e0m m\u1eabu \u0111\u1ea7u v\u00e0o c\u1ee7a ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i. Ngo\u00e0i ra, c\u1ea3 PCR v\u00e0 qPCR \u0111\u1ec1u s\u1eed d\u1ee5ng m\u00e1y lu\u00e2n nhi\u1ec7t \u0111\u1ec3 thay \u0111\u1ed5i nhi\u1ec7t \u0111\u1ed9 theo chu k\u1ef3 nh\u1eb1m th\u1ef1c hi\u1ec7n ph\u1ea3n \u1ee9ng nh\u00e2n b\u1ea3n gene.<\/p>\n\n

\u00a0<\/div>\n\n

4. S\u1ef1 kh\u00e1c nhau gi\u1eefa PCR v\u00e0 qPCR<\/strong><\/h2>\n\n

V\u1ec1 nguy\u00ean t\u1eafc, c\u1ea3 PCR v\u00e0 real-time PCR \u0111\u1ec1u d\u1ef1a tr\u00ean s\u1ef1 b\u1eaft c\u1eb7p \u0111\u1eb7c hi\u1ec7u gi\u1eefa c\u1eb7p m\u1ed3i v\u00e0 tr\u00ecnh t\u1ef1 gene m\u1ee5c ti\u00eau. Tuy nhi\u00ean, trong khi PCR ch\u1ec9 c\u00f3 th\u1ec3 coi k\u1ebft qu\u1ea3 sau khi ph\u1ea3n \u1ee9ng x\u1ea3y ra ho\u00e0n to\u00e0n th\u00ec ph\u1ea3n \u1ee9ng Real-time PCR c\u00f3 th\u1ec3 theo d\u00f5i ph\u1ea3n \u1ee9ng di\u1ec5n ra theo th\u1eddi gian th\u1ef1c. \u0110i\u1ec1u n\u00e0y c\u00f3 \u00fd ngh\u0129a quan tr\u1ecdng trong \u1ee9ng d\u1ee5ng c\u1ee7a qPCR, do n\u00f3 quy\u1ebft \u0111\u1ecbnh kh\u1ea3 n\u0103ng \u0111\u1ecbnh l\u01b0\u1ee3ng DNA c\u00f3 trong m\u1eabu.<\/p>\n\n

\u0110\u1ec3 th\u1ef1c hi\u1ec7n \u0111\u1ecbnh l\u01b0\u1ee3ng \u0111\u01b0\u1ee3c m\u1eabu, qPCR s\u1eed d\u1ee5ng thu\u1ed1c nhu\u1ed9m hu\u1ef3nh quang ho\u1eb7c m\u1eabu d\u00f2 hu\u1ef3nh quang c\u00f3 kh\u1ea3 n\u0103ng g\u1eafn v\u1edbi tr\u00ecnh t\u1ef1 DNA m\u1ee5c ti\u00eau. V\u1ec1 c\u01a1 ch\u1ebf, trong ph\u1ea3n \u1ee9ng real-time PCR, c\u00e1c ph\u00e2n t\u1eed hu\u1ef3nh quang g\u1eafn v\u1edbi DNA m\u1ee5c ti\u00eau s\u1ebd \u0111\u01b0\u1ee3c ho\u1ea1t h\u00f3a b\u1edfi tia laser v\u00e0 k\u00edch th\u00edch ch\u00fang ph\u00e1t ra \u00e1nh s\u00e1ng, sau \u0111\u00f3 \u00e1nh s\u00e1ng n\u00e0y s\u1ebd \u0111\u01b0\u1ee3c thu nh\u1eadn b\u1edfi c\u00e1c detector c\u1ee7a m\u00e1y lu\u00e2n nhi\u1ec7t chuy\u00ean cho qPCR. B\u1eb1ng c\u00e1ch \u0111o c\u01b0\u1eddng \u0111\u1ed9 t\u00edn hi\u1ec7u hu\u1ef3nh quang thu nh\u1eadn \u0111\u01b0\u1ee3c theo th\u1eddi gian di\u1ec5n ra ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i, h\u1ec7 th\u1ed1ng x\u1eed l\u00fd t\u00edn hi\u1ec7u s\u1ebd tr\u1ea3 ra \u0111\u1ed3ng th\u1eddi k\u1ebft qu\u1ea3 ph\u00e1t hi\u1ec7n t\u00e1c nh\u00e2n m\u1ee5c ti\u00eau (\u0111\u1ecbnh t\u00ednh) c\u0169ng nh\u01b0 \u0111\u1ecbnh l\u01b0\u1ee3ng DNA c\u00f3 trong m\u1eabu ban \u0111\u1ea7u (\u0111\u1ecbnh l\u01b0\u1ee3ng).\u00a0<\/p>\n\n

V\u1ec1 c\u00e1ch th\u1ee9c \u0111\u1ecdc k\u1ebft qu\u1ea3, ng\u01b0\u1eddi d\u00f9ng k\u1ef9 thu\u1eadt PCR ch\u1ec9 c\u00f3 th\u1ec3 bi\u1ebft \u0111\u01b0\u1ee3c sau khi ph\u1ea3n \u1ee9ng k\u1ebft th\u00fac. L\u00fac n\u00e0y k\u1ef9 thu\u1eadt \u0111i\u1ec7n di \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 coi ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i c\u00f3 ph\u00e1t hi\u1ec7n \u0111\u01b0\u1ee3c tr\u00ecnh t\u1ef1 m\u1ee5c ti\u00eau hay kh\u00f4ng. S\u1ea3n ph\u1ea9m PCR s\u1ebd \u0111\u01b0\u1ee3c quan s\u00e1t trong gel agarose d\u01b0\u1edbi t\u00e1c d\u1ee5ng c\u1ee7a d\u00f2ng \u0111i\u1ec7n v\u00e0 \u00e1nh s\u00e1ng UV. C\u1ee5 th\u1ec3, DNA t\u00edch \u0111i\u1ec7n \u00e2m s\u1ebd di chuy\u1ec3n v\u1ec1 ph\u00eda c\u1ef1c d\u01b0\u01a1ng trong gel d\u01b0\u1edbi t\u00e1c \u0111\u1ed9ng c\u1ee7a \u0111i\u1ec7n tr\u01b0\u1eddng, l\u00fac n\u00e0y ph\u00e2n t\u1eed DNA s\u1ebd \u0111\u01b0\u1ee3c ph\u00e2n t\u00e1ch theo kh\u1ed1i l\u01b0\u1ee3ng c\u1ee7a ch\u00fang. Gel v\u1edbi n\u1ed3ng \u0111\u1ed9 agarose cao s\u1ebd c\u00f3 k\u00edch th\u01b0\u1edbc l\u1ed7 nh\u1ecf v\u00e0 \u0111\u01b0\u1ee3c d\u00f9ng ph\u00e2n t\u00e1ch c\u00e1c ph\u00e2n t\u1eed c\u00f3 kh\u1ed1i l\u01b0\u1ee3ng nh\u1ecf, trong khi gel c\u00f3 n\u1ed3ng \u0111\u1ed9 agarose th\u1ea5p s\u1ebd ph\u00e2n t\u00e1ch \u0111\u01b0\u1ee3c c\u00e1c ph\u00e2n t\u1eed c\u00f3 kh\u1ed1i l\u01b0\u1ee3ng l\u1edbn h\u01a1n do k\u00edch th\u01b0\u1edbc l\u1ed5 l\u1edbn.<\/p>\n\n

B\u00ean c\u1ea1nh \u0111\u00f3, thang DNA (DNA ladder) \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng l\u00e0 gi\u00e1 tr\u1ecb tham chi\u1ebfu x\u00e1c \u0111\u1ecbnh k\u00edch th\u01b0\u1edbc c\u1ee7a DNA m\u1ee5c ti\u00eau. Sau khi \u0111i\u1ec7n di, b\u1ea3n gel ch\u1ee9a DNA s\u1ebd \u0111\u01b0\u1ee3c nhu\u1ed9m v\u1edbi c\u00e1c thu\u1ed1c nhu\u1ed9m (GelRed hay EtBr) v\u00e0 \u0111\u01b0a v\u00e0o m\u00e1y \u0111\u1ecdc gel ho\u1eb7c b\u00e0n soi tia UV trong bu\u1ed3ng t\u1ed1i \u0111\u1ec3 quan s\u00e1t h\u00ecnh \u1ea3nh v\u1ea1ch DNA tr\u00ean b\u1ea3n gel. D\u1ef1a v\u00e0o k\u1ebft qu\u1ea3 ghi nh\u1eadn sau \u0111i\u1ec7n di s\u1ebd bi\u1ebft \u0111\u01b0\u1ee3c x\u00e9t nghi\u1ec7m PCR c\u00f3 ph\u00e1t hi\u1ec7n \u0111\u01b0\u1ee3c t\u00e1c nh\u00e2n m\u1ee5c ti\u00eau hay kh\u00f4ng.<\/p>\n\n

Nh\u01b0 v\u1eady n\u1ebfu s\u1eed d\u1ee5ng PCR th\u00ec c\u1ea7n th\u1ef1c hi\u1ec7n th\u00eam \u0111i\u1ec7n di \u0111\u1ec3 \u0111\u1ecdc \u0111\u01b0\u1ee3c k\u1ebft qu\u1ea3 x\u00e9t nghi\u1ec7m. Trong khi \u0111\u00f3, \u0111\u1ed1i v\u1edbi real-time PCR, vi\u1ec7c \u0111\u1ecdc k\u1ebft qu\u1ea3 \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n th\u00f4ng qua quan s\u00e1t bi\u1ec3u \u0111\u1ed3 c\u01b0\u1eddng \u0111\u1ed9 t\u00edn hi\u1ec7u hu\u1ef3nh quang thu nh\u1eadn \u0111\u01b0\u1ee3c khi ph\u1ea3n \u1ee9ng di\u1ec5n ra. Hi\u1ec7n nay c\u00e1c m\u00e1y lu\u00e2n nhi\u1ec7t real-time PCR \u0111\u1ec1u k\u1ebft n\u1ed1i v\u1edbi m\u00e1y vi t\u00ednh (ho\u1eb7c t\u00edch h\u1ee3p chung) \u0111\u1ec3 \u0111\u1ecdc k\u1ebft qu\u1ea3 ph\u1ea3n \u1ee9ng. \u0110i\u1ec1u \u0111\u00f3 cho th\u1ea5y real-time PCR t\u1ed1i \u01b0u h\u01a1n PCR v\u1ec1 t\u1ed5ng th\u1eddi gian th\u1ef1c hi\u1ec7n, c\u1ee5 th\u1ec3 \u0111\u1ed1i v\u1edbi PCR s\u1ebd c\u1ea7n 3 \u0111\u1ebfn 4 gi\u1edd \u0111\u1ec3 chu\u1ea9n b\u1ecb, th\u1eddi gian di\u1ec5n ra ph\u1ea3n \u1ee9ng nh\u00e2n b\u1ea3n v\u00e0 \u0111i\u1ec7n di trong khi qPCR ch\u1ec9 m\u1ea5t kho\u1ea3ng 1,5 \u0111\u1ebfn 2 gi\u1edd \u0111\u1ec3 ho\u00e0n th\u00e0nh. Nh\u01b0 v\u1eady b\u00ean c\u1ea1nh c\u00e1c \u0111i\u1ec3m t\u01b0\u01a1ng \u0111\u1ed3ng, hai k\u1ef9 thu\u1eadt n\u00e0y c\u0169ng c\u00f3 c\u00e1c \u0111i\u1ec3m kh\u00e1c nhau ri\u00eang bi\u1ec7t \u0111\u01b0\u1ee3c t\u00f3m t\u1eaft trong B\u1ea3ng 1.<\/p>\n\n

B\u1ea3ng 1. \u0110i\u1ec3m kh\u00e1c nhau gi\u1eefa PCR v\u00e0 Real-time PCR<\/em><\/strong><\/p>\n\n

\n\n\n\n\n\n\n\n
\u00a0<\/td>\nPCR<\/strong><\/td>\nReal-time PCR<\/strong><\/td>\n<\/tr>\n
\u0110\u1ecbnh ngh\u0129a<\/strong><\/td>\nPh\u1ea3n \u1ee9ng ph\u00e1t hi\u1ec7n v\u00e0 nh\u00e2n b\u1ea3n DNA m\u1ee5c ti\u00eau c\u00f3 trong m\u1eabu (\u0111\u1ecbnh t\u00ednh)<\/td>\nPh\u1ea3n \u1ee9ng ph\u00e1t hi\u1ec7n, nh\u00e2n b\u1ea3n (\u0111\u1ecbnh t\u00ednh) v\u00e0 x\u00e1c \u0111\u1ecbnh n\u1ed3ng \u0111\u1ed9 DNA m\u1ee5c ti\u00eau c\u00f3 trong m\u1eabu (\u0111\u1ecbnh l\u01b0\u1ee3ng)<\/td>\n<\/tr>\n
Th\u00e0nh ph\u1ea7n ph\u1ea3n \u1ee9ng<\/strong><\/td>\nC\u1eb7p m\u1ed3i \u0111\u1eb7c hi\u1ec7u (primer), DNA khu\u00f4n (template DNA ), DNA polymerase, PCR buffer<\/td>\nM\u1eabu d\u00f2 (probe)\/thu\u1ed1c nhu\u1ed9m hu\u1ef3nh quang, c\u1eb7p m\u1ed3i \u0111\u1eb7c hi\u1ec7u (primer), DNA khu\u00f4n (template DNA ), DNA polymerase, PCR buffer<\/td>\n<\/tr>\n
\u0110\u1ecdc k\u1ebft qu\u1ea3 x\u00e9t nghi\u1ec7m<\/strong><\/td>\nSau khi k\u1ebft th\u00fac ph\u1ea3n \u1ee9ng, s\u1eed d\u1ee5ng k\u1ef9 thu\u1eadt \u0111i\u1ec7n di \u0111\u1ec3 xem v\u1ea1ch DNA tr\u00ean b\u1ea3n gel\u00a0<\/td>\nTheo d\u00f5i k\u1ebft qu\u1ea3 trong th\u1eddi gian th\u1ef1c t\u1ebf di\u1ec5n ra ph\u1ea3n \u1ee9ng. Xem k\u1ebft qu\u1ea3 t\u1eeb bi\u1ec3u \u0111\u1ed3 bi\u1ec3u di\u1ec5n \u0111\u01b0\u1eddng c\u01b0\u1eddng \u0111\u1ed9 t\u00edn hi\u1ec7u hu\u1ef3nh quang tr\u00ean m\u00e1y t\u00ednh<\/td>\n<\/tr>\n
Th\u1eddi gian th\u1ef1c hi\u1ec7n to\u00e0n b\u1ed9 x\u00e9t nghi\u1ec7m<\/strong><\/td>\nTrong kho\u1ea3ng 3-4 gi\u1edd bao g\u1ed3m: chu\u1ea9n b\u1ecb ph\u1ea3n \u1ee9ng, th\u1ef1c hi\u1ec7n ph\u1ea3n \u1ee9ng v\u00e0 \u0111i\u1ec7n di s\u1ea3n ph\u1ea9m PCR \u0111\u1ec3 \u0111\u1ecdc k\u1ebft qu\u1ea3 tr\u00ean b\u1ea3n gel\u00a0<\/td>\nTrong kho\u1ea3ng 1,5 -2 gi\u1edd bao g\u1ed3m: chu\u1ea9n b\u1ecb ph\u1ea3n \u1ee9ng, th\u1ef1c hi\u1ec7n ph\u1ea3n \u1ee9ng v\u00e0 \u0111\u1ecdc k\u1ebft qu\u1ea3 ngay tr\u00ean m\u00e1y t\u00ednh<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/figure>\n\n
\u00a0<\/div>\n\n

5. \u0110\u1eb7c \u0111i\u1ec3m n\u1ed5i tr\u1ed9i c\u1ee7a Real-time PCR<\/strong><\/h2>\n\n

Nh\u01b0 v\u1eady, nh\u00ecn chung Real-time PCR kh\u00e1c bi\u1ec7t so v\u1edbi PCR truy\u1ec1n th\u1ed1ng \u1edf ch\u1ed7 k\u1ef9 thu\u1eadt n\u00e0y cho ph\u00e9p theo d\u00f5i s\u1ef1 gia t\u0103ng b\u1ea3n sao DNA m\u1ee5c ti\u00eau qua m\u1ed7i chu k\u1ef3 nhi\u1ec7t \u0111\u1ed9. S\u1ef1 t\u00ednh to\u00e1n v\u00e0 ph\u00e2n t\u00edch d\u1eef li\u1ec7u t\u1eeb c\u00e1c t\u00edn hi\u1ec7u hu\u1ef3nh quang cung c\u1ea5p th\u00f4ng tin v\u1ec1 l\u01b0\u1ee3ng DNA m\u1ee5c ti\u00eau nucleic acid ban \u0111\u1ea7u trong m\u1eabu. \u0110i\u1ec1u n\u00e0y t\u1ea1o ra \u0111i\u1ec3m n\u1ed5i b\u1eadt c\u1ee7a Real-time PCR so v\u1edbi c\u00e1c x\u00e9t nghi\u1ec7m kh\u00e1c \u0111\u00f3 l\u00e0 cho ph\u00e9p ph\u00e1t hi\u1ec7n v\u00e0 x\u00e1c \u0111\u1ecbnh n\u1ed3ng \u0111\u1ed9 DNA m\u1ee5c ti\u00eau c\u00f3 trong m\u1eabu m\u1ed9t c\u00e1ch ch\u00ednh x\u00e1c (\u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u cao), \u0111\u1ed3ng th\u1eddi cung c\u1ea5p k\u1ebft qu\u1ea3 nhanh ch\u00f3ng ngay khi ph\u1ea3n \u1ee9ng \u0111ang di\u1ec5n ra.<\/p>\n\n

M\u1ed9t \u01b0u \u0111i\u1ec3m n\u1ed5i b\u1eadt kh\u00e1c c\u1ee7a Real-time PCR l\u00e0 kh\u1ea3 n\u0103ng ph\u00e1t hi\u1ec7n v\u00e0 \u0111o l\u01b0\u1eddng DNA m\u1ee5c ti\u00eau trong m\u1eabu \u1edf n\u1ed3ng \u0111\u1ed9 th\u1ea5p (\u0111\u1ed9 nh\u1ea1y cao), \u0111\u1ed3ng th\u1eddi c\u0169ng c\u00f3 th\u1ec3 theo d\u00f5i \u0111\u01b0\u1ee3c s\u1ef1 bi\u1ebfn \u0111\u1ed5i c\u1ee7a ch\u00fang theo th\u1eddi gian. Nh\u1eefng \u0111i\u1ec1u n\u00e0y l\u00e0m cho k\u1ef9 thu\u1eadt Real-time PCR tr\u1edf th\u00e0nh m\u1ed9t c\u00f4ng c\u1ee5 quan tr\u1ecdng trong nghi\u00ean c\u1ee9u bi\u1ec3u hi\u1ec7n gene, ph\u00e1t hi\u1ec7n v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng c\u00e1c t\u00e1c nh\u00e2n g\u00e2y b\u1ec7nh truy\u1ec1n nhi\u1ec5m, ph\u00e1t hi\u1ec7n m\u1ea7m b\u1ec7nh trong th\u1ef1c ph\u1ea9m hay ph\u00e1t hi\u1ec7n v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng \u0111\u1ed9t bi\u1ebfn.<\/p>\n\n

\u00a0<\/div>\n\n

6. \u1ee8ng d\u1ee5ng c\u1ee7a qPCR<\/strong><\/h2>\n\n

Real-time PCR ng\u00e0y nay \u0111\u00e3 tr\u1edf th\u00e0nh m\u1ed9t k\u1ef9 thu\u1eadt \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng nhi\u1ec1u trong l\u0129nh v\u1ef1c ch\u1ea9n \u0111o\u00e1n ph\u00e2n t\u1eed v\u00e0 nghi\u00ean c\u1ee9u bi\u1ec3u hi\u1ec7n gene. Theo \u0111\u00f3, trong ch\u1ea9n \u0111o\u00e1n ph\u00e2n t\u1eed, k\u1ef9 thu\u1eadt n\u00e0y l\u00e0 c\u00f4ng c\u1ee5 \u0111\u1ec3 ph\u00e1t hi\u1ec7n v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng gene li\u00ean quan \u0111\u1ebfn b\u1ec7nh t\u1eadt t\u1eeb virus, vi khu\u1ea9n, k\u00fd sinh tr\u00f9ng,\u2026 tr\u00ean c\u1ea3 cho ng\u01b0\u1eddi, th\u00fa y, th\u1ee7y s\u1ea3n, c\u00e2y tr\u1ed3ng, th\u1ef1c ph\u1ea9m,\u2026 \u0110i\u1ec1u n\u00e0y gi\u00fap \u0111em l\u1ea1i nh\u1eefng l\u1ee3i \u00edch l\u1edbn trong vi\u1ec7c ch\u1ea9n \u0111o\u00e1n, gi\u00e1m s\u00e1t v\u00e0 ph\u00f2ng ng\u1eeba b\u1ec7nh t\u1eadt, \u0111\u1ed3ng th\u1eddi cung c\u1ea5p th\u00f4ng tin quan tr\u1ecdng cho nghi\u00ean c\u1ee9u v\u00e0 ph\u00e2n t\u00edch d\u1ecbch t\u1ec5 h\u1ecdc.\u00a0<\/p>\n\n

\u00a0<\/div>\n\n

7. Vai tr\u00f2 c\u1ee7a ch\u1ea9n \u0111o\u00e1n Sinh h\u1ecdc ph\u00e2n t\u1eed<\/strong><\/h2>\n\n
\u00a0<\/div>\n\n

Ch\u1ea9n \u0111o\u00e1n Sinh h\u1ecdc ph\u00e2n t\u1eed trong l\u0129nh v\u1ef1c Y t\u1ebf<\/em><\/strong><\/h3>\n\n

V\u1ec1 m\u1ea3ng \u1ee9ng d\u1ee5ng ch\u1ea9n \u0111o\u00e1n b\u1ec7nh tr\u00ean ng\u01b0\u1eddi, k\u1ef9 thu\u1eadt n\u00e0y \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng r\u1ed9ng r\u00e3i trong ph\u00e1t hi\u1ec7n v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng gene t\u1eeb virus g\u00e2y b\u1ec7nh nh\u01b0 HBV, HCV, SARS-CoV-2, \u2026 Nhi\u1ec1u nghi\u00ean c\u1ee9u \u0111\u00e3 s\u1eed d\u1ee5ng Real-time PCR \u0111\u1ec3 \u0111\u1ecbnh l\u01b0\u1ee3ng gene HBV, HCV trong m\u1eabu m\u00e1u t\u1eeb b\u1ec7nh nh\u00e2n nhi\u1ec5m vi\u00eam gan (Halfon v\u00e0 c\u1ed9ng s\u1ef1, 2006; Vermehren v\u00e0 c\u1ed9ng s\u1ef1, 2008; Kania v\u00e0 c\u1ed9ng s\u1ef1, 2014)<\/em>. Nh\u1eefng nghi\u00ean c\u1ee9u n\u00e0y nh\u1ea5n m\u1ea1nh v\u00e0o vi\u1ec7c s\u1eed d\u1ee5ng Real-time PCR nh\u01b0 m\u1ed9t c\u00f4ng c\u1ee5 quan tr\u1ecdng trong ch\u1ea9n \u0111o\u00e1n vi\u00eam gan v\u00e0 \u0111\u00e1nh gi\u00e1 t\u00ecnh tr\u1ea1ng l\u00e2y nhi\u1ec5m.<\/p>\n\n

Ch\u00ednh v\u00ec v\u1eady, hi\u1ec7n nay k\u1ef9 thu\u1eadt n\u00e0y \u0111\u00e3 \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng ph\u00e1t tri\u1ec3n th\u00e0nh c\u00e1c b\u1ed9 kit \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng r\u1ed9ng r\u00e3i tr\u00ean th\u1ecb tr\u01b0\u1eddng. C\u00e1c b\u1ed9 kit n\u00e0y cung c\u1ea5p c\u00e1c c\u00f4ng c\u1ee5 c\u1ea7n thi\u1ebft \u0111\u1ec3 th\u1ef1c hi\u1ec7n quy tr\u00ecnh Real-time PCR nh\u1eb1m ph\u00e1t hi\u1ec7n v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng gene c\u1ee7a virus HBV, HCV trong m\u1eabu m\u00e1u ho\u1eb7c m\u1eabu m\u00f4 kh\u00e1c trong ch\u1ea9n \u0111o\u00e1n, qu\u00e1 tr\u00ecnh \u0111i\u1ec1u tr\u1ecb c\u0169ng nh\u01b0 \u0111\u00e1nh gi\u00e1 t\u00ecnh tr\u1ea1ng l\u00e2y nhi\u1ec5m c\u1ee7a b\u1ec7nh.<\/p>\n\n

M\u1ed9t v\u00ed d\u1ee5 kh\u00e1c \u0111\u00f3 l\u00e0 s\u1eed d\u1ee5ng Real-time PCR trong ph\u00e1t hi\u1ec7n v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng SARS-CoV-2. Nhi\u1ec1u c\u00f4ng b\u1ed1 khoa h\u1ecdc \u0111\u00e3 \u0111\u01b0\u1ee3c \u0111\u01b0a ra trong th\u1eddi \u0111i\u1ec3m di\u1ec5n ra d\u1ecbch b\u1ec7nh, trong \u0111\u00f3 real-time PCR \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng nh\u01b0 m\u1ed9t c\u00f4ng c\u1ee5 hi\u1ec7u qu\u1ea3 trong nghi\u00ean c\u1ee9u v\u00e0 \u1ee9ng d\u1ee5ng nh\u1eb1m ng\u0103n ch\u1eb7n d\u1ecbch b\u1ec7nh n\u00e0y (Park v\u00e0 c\u1ed9ng s\u1ef1, 2020; Chung v\u00e0 c\u1ed9ng s\u1ef1, 2021; Panchali v\u00e0 c\u1ed9ng s\u1ef1, 2022).<\/em> \u0110i\u1ec1u n\u00e0y cho th\u1ea5y Real-time PCR \u0111\u00f3ng vai tr\u00f2 quan tr\u1ecdng trong vi\u1ec7c x\u00e1c \u0111\u1ecbnh c\u00e1c ca nhi\u1ec5m v\u00e0 gi\u00e1m s\u00e1t t\u1ef7 l\u1ec7 l\u00e2y nhi\u1ec5m trong c\u1ed9ng \u0111\u1ed3ng.<\/p>\n\n

\u00a0<\/div>\n\n

Ch\u1ea9n \u0111o\u00e1n Sinh h\u1ecdc ph\u00e2n t\u1eed trong l\u0129nh v\u1ef1c Th\u1ee7y s\u1ea3n, Th\u00fa y, GMO<\/em><\/strong><\/h3>\n\n

\u0110\u1ed1i v\u1edbi ch\u1ea9n \u0111o\u00e1n b\u1ec7nh tr\u00ean th\u1ee7y s\u1ea3n, th\u00fa y, tr\u01b0\u1edbc \u0111\u00e2y ng\u01b0\u1eddi d\u00e2n hay ki\u1ec3m tra b\u1eb1ng c\u1ea3m quan nh\u01b0 c\u00e1c tri\u1ec7u ch\u1ee9ng b\u00ean ngo\u00e0i, nhi\u1ec7t \u0111\u1ed9 c\u01a1 th\u1ec3, theo d\u00f5i s\u1ee9c \u0103n, ho\u1ea1t \u0111\u1ed9ng c\u1ee7a v\u1eadt nu\u00f4i, ch\u1ea5t l\u01b0\u1ee3ng m\u00f4i tr\u01b0\u1eddng nu\u00f4i,\u2026 Tuy nhi\u00ean c\u00e1c ph\u01b0\u01a1ng ph\u00e1p n\u00e0y th\u01b0\u1eddng kh\u00f4ng ch\u00ednh x\u00e1c v\u00e0 kh\u00f3 ph\u00e1t hi\u1ec7n s\u1edbm c\u00e1c m\u1ea7m b\u1ec7nh. \u0110\u1ec3 h\u1ea1n ch\u1ebf thi\u1ec7t h\u1ea1i, vi\u1ec7c \u1ee9ng d\u1ee5ng c\u00f4ng ngh\u1ec7 cao trong ch\u1ea9n \u0111o\u00e1n, gi\u00e1m s\u00e1t d\u1ecbch b\u1ec7nh l\u00e0 r\u1ea5t c\u1ea7n thi\u1ebft. C\u00f9ng v\u1edbi s\u1ef1 ph\u00e1t tri\u1ec3n c\u1ee7a k\u1ef9 thu\u1eadt Real-time PCR, c\u00e1c d\u00f2ng kit \u1ee9ng d\u1ee5ng k\u1ef9 thu\u1eadt n\u00e0y \u0111ang \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng ph\u1ed5 bi\u1ebfn trong ch\u1ea9n \u0111o\u00e1n v\u00e0 theo d\u00f5i hi\u1ec7u qu\u1ea3 \u0111i\u1ec1u tr\u1ecb b\u1ec7nh.<\/p>\n\n

V\u1edbi l\u0129nh v\u1ef1c th\u1ee7y s\u1ea3n, kit Real-time PCR c\u0169ng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng nh\u1eb1m ph\u00e1t hi\u1ec7n k\u1ecbp th\u1eddi c\u00e1c t\u00e1c nh\u00e2n g\u00e2y b\u1ec7nh ph\u1ed5 bi\u1ebfn trong c\u00e1c ao nu\u00f4i nh\u01b0 virus white spot syndrome (WSSV)<\/a><\/strong> g\u00e2y b\u1ec7nh \u0111\u1ed1m tr\u1eafng \u1edf t\u00f4m, virus betanodavirus (VNN) g\u00e2y b\u1ec7nh ho\u1ea1i t\u1eed th\u1ea7n kinh \u1edf c\u00e1. Ngo\u00e0i ra, nhi\u1ec1u nh\u00e0 s\u1ea3n xu\u1ea5t \u0111\u00e3 cung c\u1ea5p nhi\u1ec1u d\u00f2ng s\u1ea3n ph\u1ea9m kit ch\u1ea9n \u0111o\u00e1n \u1ee9ng d\u1ee5ng Multiplex Real-time PCR <\/a><\/strong>nh\u1eb1m ph\u00e1t hi\u1ec7n \u0111\u1ed3ng th\u1eddi nhi\u1ec1u b\u1ec7nh tr\u00ean t\u00f4m nh\u01b0 b\u1ec7nh \u0111\u1ecf th\u00e2n t\u00f4m s\u00fa v\u00e0 \u0111\u1ecf d\u1ecdc th\u00e2n \u1ea5u tr\u00f9ng t\u00f4m s\u00fa t\u01b0\u01a1ng \u1ee9ng do vi khu\u1ea9n Vibrio vulnificus<\/em> and Vibrio alginolyticus <\/em>g\u00e2y ra. Nh\u01b0 v\u1eady kit real-time PCR \u0111\u01b0\u1ee3c d\u00f9ng \u0111\u1ec3 ph\u00e1t hi\u1ec7n t\u00e1c nh\u00e2n g\u00e2y ra tr\u00ean v\u1eadt nu\u00f4i c\u00f3 nguy c\u01a1 t\u1eed vong cao ho\u1eb7c n\u1eb1m trong y\u00eau c\u1ea7u x\u00e9t nghi\u1ec7m \u0111\u1ec3 \u0111\u1ee7 \u0111i\u1ec1u ki\u1ec7n xu\u1ea5t nh\u1eadp kh\u1ea9u qua c\u00e1c th\u1ecb tr\u01b0\u1eddng n\u01b0\u1edbc ngo\u00e0i.<\/p>\n\n

Theo \u0111\u00f3, c\u00e1c b\u1ed9 kit Real-time PCR ph\u00e1t hi\u1ec7n c\u00e1c t\u00e1c nh\u00e2n th\u01b0\u1eddng g\u1eb7p trong ch\u0103n nu\u00f4i nh\u01b0 African Swine Fever Virus (ASFV)<\/strong><\/a> g\u00e2y b\u1ec7nh t\u1ea3 l\u1ee3n ch\u00e2u Phi, virus g\u00e2y b\u1ec7nh c\u00fam A\/H5N1 \u1edf gia c\u1ea7m<\/strong><\/a>, \u1edf m\u1ea3ng th\u00fa c\u01b0ng c\u0169ng ph\u00e1t hi\u1ec7n c\u00e1c t\u00e1c nh\u00e2n nh\u01b0 Canine distemper virus (CDV)<\/a><\/strong> g\u00e2y b\u1ec7nh s\u00e0i s\u1ed1t (care) hay Ehrlichia<\/a><\/strong> g\u00e2y b\u1ec7nh k\u00fd sinh tr\u00f9ng m\u00e1u \u1edf ch\u00f3, m\u00e8o.<\/p>\n\n

Ngo\u00e0i ch\u1ea9n \u0111o\u00e1n sinh h\u1ecdc ph\u00e2n t\u1eed, Real-time PCR c\u00f2n \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng hi\u1ec7u qu\u1ea3 trong d\u00e1n nh\u00e3n v\u00e0 x\u00e1c \u0111\u1ecbnh h\u00e0m l\u01b0\u1ee3ng GMO. M\u1ed7i t\u1ed5 ch\u1ee9c hay qu\u1ed1c gia s\u1ebd quy \u0111\u1ecbnh ph\u1ea7n tr\u0103m GMO \u0111\u1ec3 d\u00e1n nh\u00e3n cho m\u1eabu, do \u0111\u00f3 \u0111\u1ec3 gi\u00fap doanh nghi\u1ec7p th\u1ef1c ph\u1ea9m c\u00f3 th\u1ec3 xu\u1ea5t kh\u1ea9u v\u00e0o c\u00e1c th\u1ecb tr\u01b0\u1eddng tr\u00ean th\u1ebf gi\u1edbi, vi\u1ec7c ph\u00e1t hi\u1ec7n sinh v\u1eadt bi\u1ebfn \u0111\u1ed5i gene l\u00e0 r\u1ea5t quan tr\u1ecdng.<\/p>\n\n

\u00a0<\/div>\n\n

8. Bi\u1ec3u hi\u1ec7n gene<\/strong><\/h2>\n\n

Trong nghi\u00ean c\u1ee9u, ph\u00e2n t\u00edch bi\u1ec3u hi\u1ec7n gene ngh\u0129a l\u00e0 x\u00e1c \u0111\u1ecbnh m\u1ee9c \u0111\u1ed9 ho\u1ea1t \u0111\u1ed9ng gene m\u1ee5c ti\u00eau trong c\u00e1c \u0111i\u1ec1u ki\u1ec7n kh\u00e1c nhau. \u0110i\u1ec1u n\u00e0y gi\u00fap l\u00e0m r\u00f5 vai tr\u00f2 c\u1ee7a c\u00e1c gene trong qu\u00e1 tr\u00ecnh ph\u00e1t tri\u1ec3n sinh l\u00fd v\u00e0 b\u1ec7nh l\u00fd c\u1ee7a sinh v\u1eadt, \u0111\u1ed3ng th\u1eddi hi\u1ec3u h\u01a1n v\u1ec1 c\u01a1 ch\u1ebf ho\u1ea1t \u0111\u1ed9ng c\u1ee7a gene, t\u1eeb \u0111\u00f3 \u1ee9ng d\u1ee5ng trong \u0111\u00e1nh gi\u00e1 hi\u1ec7u qu\u1ea3 \u0111i\u1ec1u tr\u1ecb d\u1ef1a tr\u00ean thay \u0111\u1ed5i bi\u1ec3u hi\u1ec7n gene, v\u00e0 c\u01a1 ch\u1ebf t\u00e1c \u0111\u1ed9ng c\u1ee7a thu\u1ed1c.<\/p>\n\n

Trong ph\u00e2n t\u00edch bi\u1ec3u hi\u1ec7n gene, Real-time PCR \u0111\u01b0\u1ee3c coi l\u00e0 c\u00f4ng c\u1ee5 quan tr\u1ecdng \u0111\u1ec3 \u0111o l\u01b0\u1eddng \u0111\u01b0\u1ee3c m\u1ee9c \u0111\u1ed9 bi\u1ec3u hi\u1ec7n c\u1ee7a gene. Nhi\u1ec1u c\u00f4ng b\u1ed1 v\u1ec1 ung th\u01b0 \u0111\u00e3 th\u1ef1c hi\u1ec7n ph\u00e2n t\u00edch bi\u1ec3u hi\u1ec7n gene b\u1eb1ng Real-time PCR. V\u00ed d\u1ee5, gene HER2 (Human Epidermal Growth Factor Receptor 2) l\u00e0 m\u1ed9t gene quan tr\u1ecdng li\u00ean quan \u0111\u1ebfn s\u1ef1 ph\u00e1t tri\u1ec3n v\u00e0 ti\u1ebfn tri\u1ec3n c\u1ee7a ung th\u01b0 v\u00fa (Lohrisch v\u00e0 c\u1ed9ng s\u1ef1, 2001)<\/em>. Nhi\u1ec1u c\u00f4ng b\u1ed1 \u0111\u00e3 s\u1eed d\u1ee5ng Real-time PCR \u0111\u1ec3 \u0111\u1ecbnh l\u01b0\u1ee3ng m\u1ee9c \u0111\u1ed9 bi\u1ec3u hi\u1ec7n c\u1ee7a gene HER2 trong m\u1eabu m\u00f4 ung th\u01b0 v\u00fa (Nistor v\u00e0 c\u1ed9ng s\u1ef1, 2006; Mendoza v\u00e0 c\u1ed9ng s\u1ef1, 2013; Gheni v\u00e0 c\u1ed9ng s\u1ef1, 2020)<\/em>. \u0110i\u1ec1u n\u00e0y m\u1edf ra c\u00e1nh c\u1eeda cho vi\u1ec7c ph\u00e1t tri\u1ec3n c\u00e1c ph\u01b0\u01a1ng ph\u00e1p \u0111i\u1ec1u tr\u1ecb d\u1ef1a tr\u00ean gene HER2, nh\u01b0 Herceptin (trastuzumab), m\u1ed9t lo\u1ea1i thu\u1ed1c ti\u00eam tr\u1ef1c ti\u1ebfp v\u00e0o t\u1ebf b\u00e0o ung th\u01b0 v\u00fa c\u00f3 bi\u1ec3u hi\u1ec7n HER2 cao.<\/p>\n\n

M\u1ed9t nghi\u00ean c\u1ee9u kh\u00e1c s\u1eed d\u1ee5ng real-time PCR \u0111\u1ec3 ph\u00e2n t\u00edch bi\u1ec3u hi\u1ec7n gene ch\u1ed1ng stress trong c\u00e2y Arabidopsis trong vi\u1ec7c th\u00edch \u1ee9ng v\u1edbi \u0111i\u1ec1u ki\u1ec7n kh\u00f4 h\u1ea1n. K\u1ebft qu\u1ea3 nghi\u00ean c\u1ee9u gi\u00fap cung c\u1ea5p th\u00f4ng tin chi ti\u1ebft v\u1ec1 c\u01a1 ch\u1ebf ph\u1ea3n \u1ee9ng c\u1ee7a c\u00e2y trong \u0111i\u1ec1u ki\u1ec7n m\u00f4i tr\u01b0\u1eddng kh\u1eafc nghi\u1ec7t, ch\u1ed1ng l\u1ea1i t\u00e1c \u0111\u1ed9ng c\u1ee7a m\u00f4i tr\u01b0\u1eddng. Nghi\u00ean c\u1ee9u c\u1ee7a Zhang v\u00e0 c\u1ed9ng s\u1ef1 (2022) \u0111\u00e3 s\u1eed d\u1ee5ng Real-time PCR \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh m\u1ee9c \u0111\u1ed9 bi\u1ec3u hi\u1ec7n c\u1ee7a gene PCST1<\/em> trong Arabidopsis, \u0111\u00e2y l\u00e0 gene quan tr\u1ecdng trong h\u1ec7 th\u1ed1ng ch\u1ed1ng l\u1ea1i t\u00e1c nh\u00e2n g\u00e2y stress \u1edf lo\u00e0i c\u00e2y n\u00e0y. K\u1ebft qu\u1ea3 nghi\u00ean c\u1ee9u ghi nh\u1eadn s\u1ef1 bi\u1ec3u hi\u1ec7n qu\u00e1 m\u1ee9c c\u1ee7a PCST1<\/em> l\u00e0m t\u0103ng t\u00ednh nh\u1ea1y c\u1ea3m v\u1edbi stress NaCl v\u00e0 mannitol \u1edf c\u00e2y Arabidopsis, cho th\u1ea5y vai tr\u00f2 quan tr\u1ecdng c\u1ee7a gene n\u00e0y trong h\u1ec7 th\u1ed1ng ch\u1ed1ng stress c\u1ee7a c\u00e2y.<\/p>\n\n

\u0110i\u1ec1u n\u00e0y cho th\u1ea5y ph\u00e2n t\u00edch bi\u1ec3u hi\u1ec7n gene b\u1eb1ng qPCR \u0111\u00f3ng vai tr\u00f2 quan tr\u1ecdng trong vi\u1ec7c x\u00e1c \u0111\u1ecbnh m\u1ee9c \u0111\u1ed9 bi\u1ec3u hi\u1ec7n c\u1ee7a c\u00e1c gene kh\u00e1ng stress, gi\u00fap hi\u1ec3u r\u00f5 h\u01a1n v\u1ec1 c\u00e1ch c\u00e2y Arabidopsis \u0111\u00e1p \u1ee9ng v\u00e0 ch\u1ed1ng l\u1ea1i c\u00e1c t\u00e1c \u0111\u1ed9ng c\u1ee7a m\u00f4i tr\u01b0\u1eddng.<\/p>\n\n

\u00a0<\/div>\n\n

9. T\u00e0i li\u1ec7u tham kh\u1ea3o<\/strong><\/h2>\n\n
    \n
  1. Chung, Y. S., Lee, N. J., Woo, S. H., Kim, J. M., Kim, H. M., Jo, H. J., … & Han, M. G. (2021). Validation of real-time RT-PCR for detection of SARS-CoV-2 in the early stages of the COVID-19 outbreak in the Republic of Korea. Scientific Reports, 11(1), 14817.<\/a><\/li>\n\n
  2. Gheni, N., & Westenberg, D. (2020). Quantitative real-time PCR assay with immunohistochemical evaluation of HER2\/neu oncogene in breast cancer patients and its correlation with clinicopathological findings. Indian Journal of Pathology and Microbiology, 63(Suppl 1), S123-S128.<\/a><\/li>\n\n
  3. Halfon, P., Bourli\u00e8re, M., P\u00e9naranda, G., Khiri, H., & Ouzan, D. (2006). Real-time PCR assays for hepatitis C virus (HCV) RNA quantitation are adequate for clinical management of patients with chronic HCV infection. Journal of clinical microbiology, 44(7), 2507-2511.<\/a><\/li>\n\n
  4. Higuchi, R., Fockler, C., Dollinger, G., & Watson, R. (1993). Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Bio\/technology, 11(9), 1026-1030.<\/li>\n\n
  5. Higuchi, R., Fockler, C., Dollinger, G., & Watson, R. (1993). Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Bio\/technology, 11(9), 1026-1030.<\/li>\n\n
  6. Kania, D., Ottomani, L., Meda, N., Peries, M., Dujols, P., Bollor\u00e9, K., … & Tuaillon, E. (2014). Performance of two real-time PCR assays for hepatitis B virus DNA detection and quantitation. Journal of virological methods, 201, 24-30.<\/li>\n\n
  7. Lawrence Panchali, M. J., Oh, H. J., Lee, Y. M., Kim, C. M., Tariq, M., Seo, J. W., … & Kim, D. M. (2022). Accuracy of real-time polymerase chain reaction in COVID-19 patients. Microbiology Spectrum, 10(1), e00591-21.<\/li>\n\n
  8. Lohrisch, C., & Piccart, M. (2001). An overview of HER2. In Seminars in oncology (Vol. 28, pp. 3- Zhang, H., Zhang, K., Liu, T., Zhang, Y., Tang, Z., Dong, J., & Wang, F. (2022). The characterization and expression analysis under stress conditions of PCST1<\/em> in Arabidopsis. Plant Signaling & Behavior, 17(1), 2134675.<\/li>\n\n
  9. Mackay, I. M. (2004). Real-time PCR in the microbiology laboratory. Clinical microbiology and infection, 10(3), 190-212.<\/li>\n\n
  10. Mendoza, G., Portillo, A., & Olmos\u2011Soto, J. (2013). Accurate breast cancer diagnosis through real-time PCR her\u20112 gene quantification using immunohistochemically-identified biopsies. Oncology letters, 5(1), 295-298.<\/li>\n\n
  11. Nistor, A., Watson, P. H., Pettigrew, N., Tabiti, K., Dawson, A., & Myal, Y. (2006). Real-time PCR complements immunohistochemistry in the determination of HER-2\/neu status in breast cancer. BMC clinical pathology, 6(1), 1-8.11). WB Saunders.<\/li>\n\n
  12. Park, M., Won, J., Choi, B. Y., & Lee, C. J. (2020). Optimization of primer sets and detection protocols for SARS-CoV-2 of coronavirus disease 2019 (COVID-19) using PCR and real-time PCR. Experimental & molecular medicine, 52(6), 963-977.<\/li>\n\n
  13. Suriady, W., Putra, A. C., Wiyono, W. H., Alatas, M. F., Bermawi, B., Moniqa, R., … & Sumargono, J. E. (2020). An atypical case: RT-PCR-negative, sero-positive covid-19 patient. Pneumologia, 69(2), 107-114.<\/li>\n\n
  14. Vermehren, J., Kau, A., G\u00e4rtner, B. C., G\u00f6bel, R., Zeuzem, S., & Sarrazin, C. (2008). Differences between two real-time PCR-based hepatitis C virus (HCV) assays (RealTime HCV and Cobas AmpliPrep\/Cobas TaqMan) and one signal amplification assay (Versant HCV RNA 3.0) for RNA detection and quantification. Journal of clinical microbiology, 46(12), 3880-3891.<\/li>\n<\/ol>\n\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>","protected":false},"excerpt":{"rendered":"

    PCR (Polymerase Chain Reaction) l\u00e0 k\u1ef9 thu\u1eadt sinh h\u1ecdc ph\u00e2n t\u1eed nh\u00e2n b\u1ea3n gene m\u1ee5c ti\u00eau l\u00ean h\u00e0ng tri\u1ec7u l\u1ea7n trong m\u00f4i tr\u01b0\u1eddng in vitro d\u1ef1a v\u00e0o s\u1ef1 thay \u0111\u1ed5i nhi\u1ec7t \u0111\u1ed9 theo chu k\u1ef3, qu\u00e1 tr\u00ecnh nh\u00e2n b\u1ea3n \u0111\u01b0\u1ee3c m\u00f4 ph\u1ecfng theo c\u01a1 ch\u1ebf c\u1ee7a qu\u00e1 tr\u00ecnh sao ch\u00e9p DNA trong t\u1ebf b\u00e0o (Putra v\u00e0 c\u1ed9ng s\u1ef1, 2020). K\u1ef9 thu\u1eadt n\u00e0y c\u00f3 kh\u1ea3 n\u0103ng ph\u00e1t hi\u1ec7n ch\u00ednh x\u00e1c m\u1ee5c ti\u00eau v\u1edbi l\u01b0\u1ee3ng m\u1eabu t\u1ed1i thi\u1ec3u, m\u00e0 c\u00e1c ph\u01b0\u01a1ng ph\u00e1p kh\u00e1c nh\u01b0 nu\u00f4i c\u1ea5y ho\u1eb7c huy\u1ebft thanh h\u1ecdc kh\u00f4ng th\u1ec3 th\u1ef1c hi\u1ec7n \u0111\u01b0\u1ee3c. \u0110\u1ed9 \u0111\u1eb7c hi\u1ec7u v\u00e0 \u0111\u1ed9 nh\u1ea1y cao c\u1ee7a PCR l\u00e0 l\u00fd do khi\u1ebfn n\u00f3 \u0111\u01b0\u1ee3c coi l\u00e0 \u201cti\u00eau chu\u1ea9n v\u00e0ng\u201d trong c\u00e1c x\u00e9t nghi\u1ec7m ch\u1ea9n \u0111o\u00e1n b\u1ec7nh.<\/p>","protected":false},"author":1,"featured_media":10798,"comment_status":"open","ping_status":"open","sticky":true,"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":"set","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":[228,270,266,267],"tags":[480,481,448],"class_list":["post-9888","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-bai-viet","category-kien-thuc","category-sinh-hoc-phan-tu","category-sinh-hoc-phan-tu-bai-viet","tag-pcr","tag-real-tim-pcr","tag-sinh-hoc-phan-tu"],"uagb_featured_image_src":{"full":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR.jpg",1920,1080,false],"thumbnail":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-150x150.jpg",150,150,true],"medium":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-300x169.jpg",300,169,true],"medium_large":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-768x432.jpg",768,432,true],"large":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-1024x576.jpg",1024,576,true],"1536x1536":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-1536x864.jpg",1536,864,true],"2048x2048":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR.jpg",1920,1080,false],"trp-custom-language-flag":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-18x10.jpg",18,10,true],"woocommerce_thumbnail":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-1707x960.jpg",1707,960,true],"woocommerce_single":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR.jpg",1920,1080,false],"woocommerce_gallery_thumbnail":["https:\/\/abtvn.com\/wp-content\/uploads\/2024\/03\/ABT-blog-PCR-va-Real-time-PCR-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 (Polymerase Chain Reaction) l\u00e0 k\u1ef9 thu\u1eadt sinh h\u1ecdc ph\u00e2n t\u1eed nh\u00e2n b\u1ea3n gene m\u1ee5c ti\u00eau l\u00ean h\u00e0ng tri\u1ec7u l\u1ea7n trong m\u00f4i tr\u01b0\u1eddng in vitro d\u1ef1a v\u00e0o s\u1ef1 thay \u0111\u1ed5i nhi\u1ec7t \u0111\u1ed9 theo chu k\u1ef3, qu\u00e1 tr\u00ecnh nh\u00e2n b\u1ea3n \u0111\u01b0\u1ee3c m\u00f4 ph\u1ecfng theo c\u01a1 ch\u1ebf c\u1ee7a qu\u00e1 tr\u00ecnh sao ch\u00e9p DNA trong t\u1ebf b\u00e0o (Putra…","_links":{"self":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/9888","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=9888"}],"version-history":[{"count":17,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/9888\/revisions"}],"predecessor-version":[{"id":13747,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/9888\/revisions\/13747"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/media\/10798"}],"wp:attachment":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/media?parent=9888"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/categories?post=9888"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/tags?post=9888"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}