Ukumelana ne-Mycobacterium Tuberculosis Nucleic Acid kanye ne-Rifampicin
Igama lomkhiqizo
Ikhithi Yokuthola Ukumelana Ne-HWTS-RT074B-Mycobacterium Tuberculosis Nucleic Acid kanye ne-Rifampicin (Melting Curve)
Isitifiketi
CE
I-Epidemiology
I-Mycobacterium tuberculosis, njengoba nje i-Tubercle bacillus, i-TB, iyigciwane elibangela isifo sofuba. Njengamanje, imithi yokuqala yokulwa nesifo sofuba esetshenziswa kakhulu ihlanganisa i-isoniazid, i-rifampicin ne-hexambutol, njll. Imithi yesibili yokulwa nesifo sofuba ihlanganisa i-fluoroquinolones, i-amikacin ne-kanamycin, njll. Imithi emisha ethuthukisiwe yi-linezolid, i-bedaquiline ne-delamani, njll. Kodwa-ke, ngenxa yokusetshenziswa okungalungile kwemithi yokulwa nesifo sofuba kanye nezici zesakhiwo sodonga lweseli se-mycobacterium tuberculosis, i-mycobacterium tuberculosis ithuthukisa ukumelana nemithi yemithi yokulwa nesifo sofuba, okuletha izinselele ezinkulu ekuvimbeleni nasekwelapheni isifo sofuba.
I-Rifampicin ibilokhu isetshenziswa kabanzi ekwelapheni iziguli ezine-TB yamaphaphu kusukela ngasekupheleni kwawo-1970, futhi inethonya elikhulu. Kube yisinqumo sokuqala ukufinyeza ukwelashwa ngamakhemikhali kweziguli ezine-TB yamaphaphu. Ukumelana ne-Rifampicin kubangelwa kakhulu ukuguquka kwezakhi zofuzo ze-rpoB. Nakuba imithi emisha yokulwa nesifo sofuba iphuma njalo, futhi ukusebenza kahle kweziguli ezine-TB yamaphaphu nakho kuqhubekile nokuthuthuka, kusenokuntuleka kwemithi yokulwa nesifo sofuba, futhi isimo sokusetshenziswa kwemithi okungenangqondo emtholampilo siphezulu kakhulu. Ngokusobala, isifo sofuba se-Mycobacterium ezigulini ezine-TB yamaphaphu asikwazi ukubulawa ngokuphelele ngesikhathi esifanele, okugcina kuholele emazingeni ahlukene okumelana nemithi emzimbeni wesiguli, kwandise isikhathi sesifo, futhi kwandisa ingozi yokufa kwesiguli.
Isiteshi
| Isiteshi | Iziteshi nama-Fluorophores | I-Reaction Buffer A | I-Reaction Buffer B | I-Reaction Buffer C |
| Isiteshi se-FAM | Intatheli: FAM, Ukucima: Akukho | i-rpoB 507-514 | i-rpoB 513-520 | 38KD kanye ne-IS6110 |
| Isiteshi se-CY5 | Intatheli: CY5, i-Quencher: Akukho | i-rpoB 520-527 | i-rpoB 527-533 | / |
| Isiteshi se-HEX (VIC) | Intatheli: HEX (VIC), Quencher: Akukho | Ukulawula kwangaphakathi | Ukulawula kwangaphakathi | Ukulawula kwangaphakathi |
Amapharamitha Obuchwepheshe
| Isitoreji | ≤-18℃ Ebumnyameni |
| Isikhathi sokuphelelwa yisikhathi | Izinyanga ezingu-12 |
| Uhlobo Lwesifanekiso | Isikhwehlela |
| CV | ≤5.0% |
| I-LoD | isifo sofuba se-mycobacterium amabhaktheriya angu-50/mL uhlobo lwasendle olungamelana ne-rifampicin: 2x103amabhaktheriya/mL i-homozygous mutant: 2x103amabhaktheriya/mL |
| Ukucaciswa | Ithola isifo sofuba se-mycobacterium esiwuhlobo lwasendle kanye nezindawo zokuguquka kwezakhi zofuzo ezimelana nemithi njenge-katG 315G>C\A, InhA-15C> T, imiphumela yokuhlolwa ayikhombisi ukumelana ne-rifampicin, okusho ukuthi akukho ukusabela okuxubile. |
| Izinsimbi Ezisebenzayo: | Izinhlelo ze-PCR Zesikhathi Sangempela ze-SLAN-96P Uhlelo lwe-BioRad CFX96 Real-Time PCR Uhlelo lwe-PCR lwe-LightCycler480® lwesikhathi sangempela |
Ukugeleza Komsebenzi
Uma usebenzisa i-Macro & Micro-Test General DNA/RNA Kit (HWTS-3019-50, HWTS-3019-32, HWTS-3019-48, HWTS-3019-96) (engasetshenziswa ne-Macro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006C, HWTS-3006B)) noma i-Macro & Micro-Test Viral DNA/RNA Column (HWTS-3022-50) yi-Jiangsu Macro & Micro-Test Med-Tech Co., Ltd. ukuze kukhishwe, engeza u-200μL we-Positive Control, i-negative control kanye nesampula ye-sputum ecutshunguliwe ukuze ihlolwe ngokulandelana, bese ufaka u-10μL we-internal control ngokwehlukana ku-Positive Control, i-negative control kanye nesampula ye-sputum ecutshunguliwe ukuze ihlolwe, futhi izinyathelo ezilandelayo kufanele zenziwe ngokuqinile ngokwemiyalelo yokukhipha. Ivolumu yesampula ekhishwe ingu-200μL, kanti ivolumu yokulungisa enconywayo ingu-100μL.
