He aha ka hoʻāʻo PCR? E kamaʻilio maʻalahi kākou e pili ana!

Ke lohe kākou i ka huaʻōlelo PCR, noʻonoʻo kākou i ka maʻi ahulau Covid, ʻeā? ʻO ka manawa i pono ai mākou e hoʻokomo i kahi swab i loko o ko mākou ihu a kali i ka hōʻike e hiki mai ana... ʻaʻole loa e poina ʻia kēlā. Akā ua ʻike anei ʻoe ʻaʻole hoʻohana wale ʻia ka PCR no ka ʻike ʻana iā Covid? He hoʻāʻo koʻikoʻi loa kēia i hana i kahi holomua nui i ka ʻepekema lapaʻau. No laila i kēia lā, e kamaʻilio kākou e pili ana i ke ʻano o kēia hoʻāʻo PCR, he aha nā mea ʻē aʻe e hiki ke ʻike ʻia, a pehea e hana ai ma ke ʻano maʻalahi hiki iā ʻoe ke hoʻomaopopo.

He aha ka PCR maʻalahi?

ʻAe, e ʻōlelo kākou penei. E noʻonoʻo ʻoe e pono ʻoe e ʻimi i hoʻokahi huaʻōlelo ma kahi ʻaoʻao hoʻokahi o kahi puke hoʻokahi, mai loko mai o nā tausani o nā puke i loko o kahi hale waihona puke nui. Pehea ka paʻakikī o ia mea? Ua like ia me ka ʻimi ʻana i kahi ʻāpana liʻiliʻi o ka DNA a i ʻole RNA , ka mea genetic i loko o nā cell i loko o ko kākou kino.

ʻO ka ʻenehana PCR (Polymerase Chain Reaction) e maʻalahi ai kēlā hana paʻakikī. ʻO ka hana a kēia, ʻo ia ke kope ʻana i ka ʻāpana liʻiliʻi loa o ka DNA a i ʻole RNA a mākou e ʻimi nei no nā haneli haneli, a i ʻole miliona mau manawa (amplification). Ua like ia me ka loaʻa ʻana o ka ʻaoʻao me ka huaʻōlelo ma ka puke waihona puke, e waiho ana i loko o kahi mīkini kope, a hana i kahi miliona kope. A laila, ʻoiai ua loaʻa kēlā huaʻōlelo paʻakikī i nā wahi āpau, hiki iā mākou ke ʻike maʻalahi iā ia.

I ka ʻōlelo maʻalahi, ʻo ka PCR kahi hoʻāʻo e lawe i kahi liʻiliʻi loa o kahi cell mai ko mākou kino, kahi kulu koko, a i ʻole kahi swab ihu, a hoʻoulu hana i kahi ʻāpana DNA a RNA paha i loko o kahi keʻena hoʻokolohua e hōʻoia inā aia ia i loko o kā mākou laʻana a ʻaʻole paha. He hoʻāʻo koʻikoʻi loa kēia. ʻO ia hoʻi, hiki iā ia ke ʻike i nā liʻiliʻi loa o ka maʻi. No kēia ʻano hana kahi hui o nā hoʻāʻo i kapa ʻia ʻo Nucleic Acid Amplification Test (NAAT).

He aha nā maʻi e hiki ke ʻike ʻia e ka hoʻāʻo PCR?

ʻOiai ua lilo ka PCR i mea kaulana ma muli o ka maʻi ahulau COVID, ua hoʻohana ʻia ia i loko o ke ao lapaʻau no nā makahiki he ʻumi. Nui nā mea e hiki ke ʻike ʻia me kēia hoʻāʻo. E nānā kākou i kekahi o nā mea nui.

Pehea e lawe ai i kahi hāpana no kahi hoʻāʻo PCR?

ʻO ke ʻano o kāu lawe ʻana i kahi hāpana no kahi hoʻāʻo PCR e pili ana i ke ʻano o ka maʻi āu e ʻimi nei. He ʻokoʻa ke ʻano o ka lawe ʻana i kahi hāpana no kēlā me kēia kūlana.

ʻO ka hapa nui o ka manawa, ʻaʻole pono ʻoe e hana i kekahi mea kūikawā e hoʻomākaukau ai no kahi hoʻāʻo PCR. Eia nō naʻe, i kekahi mau hihia, e like me ka hana ʻana i kahi mea e like me ka biopsy, e hāʻawi kāu kauka iā ʻoe i nā ʻōlelo aʻoaʻo e pono ai ma mua.

Pehea e hana ai ka ʻenehana PCR?

ʻAe, e nānā kākou pehea e hana maoli ai kēia ʻenehana kupaianaha. He ʻano ʻepekema iki kēia, akā e wehewehe maʻalahi wau.

1. Hoʻomākaukau ʻana i ka laʻana: ʻO ka mea mua, ua hoʻokaʻawale ʻia ka DNA a i ʻole RNA mai ka laʻana (koko, nā mea huna, a pēlā aku) i lawe ʻia mai ou aku.

2. Ke hana ʻana i ka hui ʻana: ʻO ka mea aʻe, ua hui pū ʻia kēia DNA/RNA i hoʻokaʻawale ʻia me kekahi mau kemika kūikawā. ʻO ka mea nui i loko o kēia hui ʻana he enzyme i kapa ʻia ʻo DNA Polymerase . ʻO kēia ka mea nui e hana i ka hana kope.

3. Kaʻapuni Hoʻomehana a me ka Hoʻomaʻalili: I kēia manawa, ua waiho ʻia ka ʻōmole liʻiliʻi me kēia hui ʻana i loko o kahi mīkini kūikawā. Hoʻokiʻekiʻe a hoʻohaʻahaʻa kēia mīkini i ka mahana me ka wikiwiki loa.

• Denaturation: ʻO ka mea mua, hoʻonui ʻia ka mahana a ma kahi o 95 degere Celsius. ʻO kēia ke kumu e hoʻokaʻawale ai nā kaula DNA ʻelua.
• Hoʻoheheʻe ʻana: A laila, hoʻohaʻahaʻa ʻia ka mahana i kahi o 55 degere. A laila, hoʻopili ʻia nā ʻāpana liʻiliʻi o ka DNA (primers) e kūlike me ka ʻāpana DNA a mākou e ʻimi nei i nā kaula DNA i hoʻokaʻawale ʻia.
• Hoʻomehana hou ʻana (Hoʻonui): A laila hoʻokiʻekiʻe ʻia ka mahana i kahi o 72 degere. A laila hele mai ka enzyme DNA polymerase a hana i kahi kaula DNA hou mai kēlā primer e kūlike me ke kaula DNA mua.

4. Ke Kope ʻana: I kēia manawa, ua hana ʻia ʻelua kope o hoʻokahi kaula DNA. Hana hou ʻia kēia pōʻaiapuni mahana ma kahi o 30-40 mau manawa. I hoʻokahi pōʻaiapuni, pāpālua ka nui o ka DNA. ʻElua lilo i ʻehā, ʻehā lilo i ʻewalu, ʻewalu lilo i ʻumikumamāono... Ma hope o 30 mau pōʻaiapuni, ua hana ʻia ma mua o hoʻokahi piliona kope mai ka ʻāpana DNA mua hoʻokahi.

I kēia manawa, ʻo kēlā ʻāpana DNA/RNA, ka mea i liʻiliʻi loa no ka ʻike ʻana ma mua, ua lilo i mea nui e hiki ai ke ʻike maʻalahi ʻia.

Aia kekahi mau ʻano hoʻāʻo PCR?

ʻAe, ʻelua mau ʻano nui e pono ai mākou e makaʻala.

• ʻO ka PCR Reverse-transcriptase (RT-PCR): Loaʻa i kekahi mau virus, e like me Covid, influenza, a me HIV, kā lākou ʻike genetic i loko o ka RNA ma mua o ka DNA. Hiki i ka PCR maʻamau ke kope wale i ka DNA. No laila, ʻo ka mea āu e hana ai ma ka RT-PCR, ʻo ia ka hoʻohana mua ʻana i kahi enzyme kūikawā (`Reverse Transcriptase`) e 'hoʻololi' i kēlā RNA i DNA. ʻO ka DNA hopena ua kapa ʻia ʻo DNA complementary (cDNA). A laila hana ʻia ke kaʻina hana PCR maʻamau.

• PCR Manawa Maoli / PCR Kūʻai (Q-PCR): He ʻano hana holomua iki kēia. ʻO ka mea e hana ʻia ma kēia, ʻoiai ke kope ʻia ka DNA, ua hoʻopili ʻia kahi lepili fluorescent iā ia. I ka piʻi ʻana o ka nui o ka DNA i kope ʻia, piʻi pū kēia fluorescence. Hiki i ka mīkini ke ana i kēia fluorescence. No laila, ʻoiai mākou e kali ana, ʻo ia hoʻi, i ka manawa maoli , hiki iā mākou ke ʻike inā aia ka DNA pili i loko o ka laʻana a i ʻole. ʻAʻole wale ia, hiki iā ia ke hāʻawi i kahi manaʻo maʻamau o ka nui o ka maʻi i loko o ka laʻana mua (viral load). Hoʻohana pinepine ʻia kahi hoʻāʻo RT-PCR Manawa Maoli e hoʻohui i kēia mau ʻano ʻelua e ʻike iā Covid.

He aha ka manaʻo o ka hōʻike PCR?

He mea nui loa e hoʻomaopopo i kahi hōʻike PCR ke loaʻa iā ʻoe.

• ʻIke ʻia / Maikaʻi: Inā ʻōlelo kāu hōʻike "ʻIke ʻia" a i ʻole "Maikaʻi," ʻo ia hoʻi, aia i loko o kāu hāpana nā ʻāpana o ka maʻi a i ʻole nā ​​mea genetic a lākou e ʻimi nei. I ka pōkole, ua loaʻa paha iā ʻoe ka maʻi.

• ʻAʻole i ʻike ʻia / Maikaʻi ʻole: ʻO ke ʻano kēia ʻaʻole hiki ke loaʻa nā ʻāpana DNA/RNA pili i loko o ka hāpana i hoʻāʻo ʻia. ʻO ke ʻano kēia ʻaʻole paha ʻoe i loaʻa i kēlā maʻi.

ʻOkoʻa ka lōʻihi o ka loaʻa ʻana o nā hopena ma muli o ka hoʻāʻo e hana ʻia nei. Loaʻa pinepine nā hopena i nā swabs ihu (e like me COVID, influenza) i loko o 24 mau hola. Eia nō naʻe, hiki i nā hoʻāʻo koko a me nā biopsies e ʻimi ai i nā maʻi genetic ke hala i hoʻokahi pule a ʻoi aku paha.

Inā he mau nīnau a hopohopo paha kāu e pili ana i kahi hoʻāʻo PCR a i ʻole kona mau hopena, e kamaʻilio pono me kāu kauka e pili ana i ia mea.