Isikhali Esisha Sokuxilonga Isifo Sofuba kanye Nokuthola Ukumelana Nemithi: Ukulandelana Okuqondiwe Kwesizukulwane Esisha (tNGS) Okuhlanganiswe Nokufunda Komshini Kokuxilonga Isifo Sofuba Ngokuzwela Okuphezulu
Umbiko wezincwadi: I-CCa: imodeli yokuxilonga esekelwe ku-tNGS kanye nokufunda komshini, efanelekile kubantu abanesifo sofuba esingabangwa amagciwane kanye ne-tuberculous meningitis.
Isihloko sethisisi: Ukulandelana kwesizukulwane esilandelayo okuqondiswe ku-tuberculous kanye nokufunda komshini: isu lokuxilonga elibucayi kakhulu lama-tubular e-pulmonary aphansi kanye ne-meningitis e-tubular.
Isikhathi: 《Clinica Chimica Acta》
UMA: 6.5
Usuku lokushicilelwa: Januwari 2024
Kuhlanganiswe ne-University of Chinese Academy of Sciences kanye ne-Beijing Chest Hospital of Capital Medical University, i-Macro & Micro-Test yasungula imodeli yokuxilonga isifo sofuba esekelwe esizukulwaneni esisha sobuchwepheshe bokulandelana okuqondiwe (tNGS) kanye nendlela yokufunda komshini, enikeze ukuzwela okuphezulu kakhulu kokuthola isifo sofuba esinamabhaktheriya ambalwa kanye ne-tuberculous meningitis, yanikeza indlela entsha yokuxilonga isifo sofuba esibangelwa ukucindezeleka kokuxilongwa kwezinhlobo ezimbili zesifo sofuba, futhi yasiza ekuxilongweni okunembile, ekutholakaleni kokumelana nemithi kanye nokwelashwa kwesifo sofuba. Ngesikhathi esifanayo, kutholakale ukuthi i-plasma cfDNA yesiguli ingasetshenziswa njengohlobo lwesampula olufanele lokuthola isampula yomtholampilo ekuxilongweni kwe-TBM.
Kulolu cwaningo, amasampula e-plasma angu-227 kanye namasampula oketshezi lwe-cerebrospinal asetshenziswe ukusungula amaqembu amabili emitholampilo, lapho amasampula e-laboratory diagnostic cohort asetshenziswa ukusungula imodeli yokufunda komshini yokuxilongwa kwesifo sofuba, kanti amasampula e-clinical diagnostic cohort asetshenziswa ukuqinisekisa imodeli yokuxilonga esungulwe. Wonke amasampula aqale aqondiswa yi-tap pool ehlosiwe yokuthwebula isifo sofuba se-Mycobacterium. Ngemuva kwalokho, ngokusekelwe kudatha yokulandelana kwe-TB-tNGS, imodeli yesihlahla sesinqumo isetshenziswa ukwenza ukuqinisekiswa okuphindwe kahlanu kumasethi okuqeqeshwa nokuqinisekiswa komugqa wokuxilonga welabhorethri, futhi kutholakala imikhawulo yokuxilonga yamasampula e-plasma kanye namasampula oketshezi lwe-cerebrospinal. Umkhawulo otholiwe ulethwa kumasethi amabili okuhlolwa komugqa wokuxilonga wezokwelapha ukuze kutholakale, futhi ukusebenza kokuxilonga komfundi kuhlolwa nge-ROC curve. Ekugcineni, kutholakale imodeli yokuxilonga isifo sofuba.
Umfanekiso 1 umdwebo wesakhiwo socwaningo
Imiphumela: Ngokwemingcele ethile yesampula ye-CSF DNA (AUC = 0.974) kanye nesampula ye-plasma cfDNA (AUC = 0.908) enqunywe kulolu cwaningo, phakathi kwamasampula angu-227, ukuzwela kwesampula ye-CSF kwakungu-97.01%, ukucaciswa kwakungu-95.65%, kanti ukuzwela nokucaciswa kwesampula ye-plasma kwakungu-82.61% no-86.36%. Ekuhlaziyweni kwamasampula angu-44 abhangqiwe e-plasma cfDNA kanye ne-cerebrospinal fluid DNA evela ezigulini ze-TBM, isu lokuxilonga lalolu cwaningo linokuhambisana okuphezulu okungu-90.91% (40/44) ku-plasma cfDNA kanye ne-cerebrospinal fluid DNA, kanti ukuzwela kungama-95.45% (42/44). Ezinganeni ezinesifo sofuba samaphaphu, isu lokuxilonga lalolu cwaningo lizwela kakhulu kumasampula e-plasma kunemiphumela yokutholwa kwe-Xpert yamasampula ejusi lesisu avela ezigulini ezifanayo (28.57% VS 15.38%).
Umfanekiso 2 Ukuhlaziywa kokusebenza kwemodeli yokuxilonga isifo sofuba kumasampula abantu
Isithombe 3 Imiphumela yokuxilonga yamasampula abhangqiwe
Isiphetho: Indlela yokuxilonga isifo sofuba ezwela kakhulu yasungulwa kulolu cwaningo, enganikeza ithuluzi lokuxilonga elinozwela oluphezulu kakhulu lokubona iziguli zezokwelapha ezine-oligobacillary tuberculosis (isiko elibi). Ukuthola isifo sofuba esizwela kakhulu ngokusekelwe ku-plasma cfDNA kungaba uhlobo lwesampula olufanele lokuxilonga isifo sofuba esisebenzayo kanye ne-tuberculous meningitis (amasampula e-plasma kulula ukuwaqoqa kune-cerebrospinal fluid ezigulini ezisolwa ukuthi zinesifo sofuba sobuchopho).
Isixhumanisi sokuqala: https://www.sciencedirect.com/science/article/pii/s0009898123004990? via%3Dihub
Isingeniso esifushane semikhiqizo yochungechunge lokuthola isifo sofuba lwe-Macro & Micro-Test
Ngenxa yesimo esiyinkimbinkimbi sesampula seziguli ezinesifo sofuba kanye nezidingo ezahlukahlukene, i-Macro & Micro-Test inikeza isethi ephelele yezixazululo ze-NGS zokukhipha uketshezi kumasampula esikhwehlela, ukwakhiwa nokulandelana komtapo wezincwadi we-Qualcomm, kanye nokuhlaziywa kwedatha. Imikhiqizo ihlanganisa ukuxilongwa okusheshayo kweziguli ezinesifo sofuba, ukutholwa kokumelana nemithi yesifo sofuba, ukuthayipha isifo sofuba se-mycobacterium kanye ne-NTM, ukuxilongwa kwe-hypersensitivity kwesifo sofuba esingenayo amagciwane kanye nabantu abanesifo sofuba, njll.
Ama-serial detection kits esifo sofuba kanye nama-mycobacteria:
| Inombolo Yento | igama lomkhiqizo | okuqukethwe kokuhlolwa komkhiqizo | uhlobo lwesampula | imodeli esebenzayo |
| I-HWTS-3012 | I-ejenti yokukhipha isampula | Isetshenziswa ekwelapheni amasampula esikhwehlela ngoketshezi, ithole inombolo yerekhodi yekilasi lokuqala, i-Sutong Machinery Equipment 20230047. | isikhwehlela | |
| I-HWTS-NGS-P00021 | Ikhithi yokuthola inani le-Qualcomm yesifo sofuba esizwela kakhulu (indlela yokubamba i-probe) | Ukutholwa kokusabela okungekuhle (i-biopsy yoketshezi) kwesifo sofuba samaphaphu esingenamagciwane kanye namaqhubu obuchopho; Amasampula abantu okusolwa ukuthi batheleleke ngesifo sofuba noma i-mycobacteria engeyona i-tuberculosis ahlaziywe nge-metagenomics ejulile, kanye nolwazi lokuthola ukuthi i-mycobacteria yesifo sofuba noma engeyona i-tuberculosis ithelelekile yini kanye nolwazi oluyinhloko lokumelana nemithi lokuqala lwesifo sofuba se-mycobacterium. | Igazi elingaphandle, uketshezi lokuhlanza i-alveolar, i-hydrothorax kanye ne-ascites, isampula yokubhoboza okugxile, uketshezi lwe-cerebrospinal. | Isizukulwane sesibili |
| I-HWTS-NGS-T001 | Ikhithi yokuthola ukuthayipha kwe-Mycobacterium kanye nokumelana nemithi (indlela yokulandelanisa i-multiplex amplification) | Ukuhlolwa kokuthayipha kwe-Mycobacterium, okuhlanganisa i-MTBC kanye ne-187 NTM；Ukutholwa kokumelana nemithi yesifo sofuba i-Mycobacterium kuhlanganisa imithi engu-13 kanye nezindawo ezingu-16 eziyinhloko zokuguqulwa kwezakhi zofuzo ezimelana nemithi. | I-sputum, uketshezi lokuhlanza i-alveolar, i-hydrothorax kanye ne-ascites, isampula yokubhoboza okugxile, uketshezi lomgogodla. | Ipulatifomu yesizukulwane sesibili/sesithathu |
Okuvelele: Ikhithi yokuthayipha ye-HWTS-NGS-T001 Mycobacterium kanye nokuthola ukumelana nemithi (indlela yokukhulisa okuningi)
Isingeniso somkhiqizo
Lo mkhiqizo usekelwe emithini eyinhloko yokuqala neyesibili echazwe eziqondisweni zokwelapha i-TB ze-WHO, ama-macrolide nama-aminoglycosides asetshenziswa kakhulu eziqondisweni zokwelapha ze-NTM, kanti izindawo zokumelana nemithi zihlanganisa lonke iqembu elilodwa lezindawo ezihlobene nokumelana nemithi kukhathalogi yokuguqulwa kwe-mutation ye-WHO Mycobacterium tuberculosis complex, kanye nezinye izakhi zofuzo zokumelana nemithi ezibikiwe kanye nezindawo zokuguquka ngokusho kophenyo kanye nezibalo zezincwadi ezithola amaphuzu aphezulu ekhaya nakwamanye amazwe.
Ukuhlonza ukuthayipha kusekelwe ezinhlotsheni ze-NTM ezifingqiwe eziqondisweni ze-NTM ezishicilelwe yi-Chinese Journal of Tuberculosis and Respiratory Diseases kanye nokuvumelana kochwepheshe. Ama-primer okuthayipha aklanyelwe angakhulisa, alandele futhi achaze izinhlobo ze-NTM ezingaphezu kuka-190.
Ngobuchwepheshe bokukhulisa i-PCR eqondisiwe, izakhi zofuzo zofuzo kanye nezakhi zofuzo ezingamelani nemithi ze-Mycobacterium zandiswa yi-PCR eningi, futhi kwatholakala inhlanganisela ye-amplicon yezakhi zofuzo eziqondiwe. Imikhiqizo ekhulisiwe ingakhiwa ibe yilabhulali yokulandelelana kwesizukulwane sesibili noma sesithathu, futhi wonke amapulatifomu okulandelelana kwesizukulwane sesibili nesesithathu angafakwa ku-sequencing ejulile ukuze kutholakale ulwazi lokulandelana kwezakhi zofuzo eziqondiwe. Ngokuqhathanisa nezinguquko ezaziwayo eziqukethwe kudathabheyisi yokubhekisela eyakhelwe ngaphakathi (kufaka phakathi ikhathalogi ye-WHO Mycobacterium tuberculosis complex mutation kanye nobudlelwano bayo nokumelana nemithi), izinguquko ezihlobene nokumelana nemithi noma ukuthambekela kwemithi yokulwa nesifo sofuba zanqunywa. Kuhlanganiswe nesisombululo sokwelashwa kwesampula ye-sputum esizivulekelayo se-Macro & Micro-Test, inkinga yokusebenza kahle kwe-nucleic acid amplification kwamasampula e-sputum emitholampilo (okuphindwe kayishumi kunezindlela zendabuko) yaxazululwa, ukuze ukutholwa kokulandelana kokumelana nemithi kungasetshenziswa ngqo kumasampula e-sputum emitholampilo.
Ububanzi bokutholwa komkhiqizo
34izakhi zofuzo ezihlobene nokumelana nemithi18imithi yokulwa nesifo sofuba kanye6Kutholakale izidakamizwa ze-NTM, ezimboza297izindawo zokumelana nemithi; Izinhlobo eziyishumi zesifo sofuba se-Mycobacterium kanye nezinye ezingaphezu kwalokho190izinhlobo ze-NTM zitholiwe.
Ithebula 1: Ulwazi lwezidakamizwa ezingu-18+6 +190+NTM
Inzuzo yomkhiqizo
Ukuzivumelanisa nezimo okunamandla kwezokwelapha: amasampula e-sputum angatholakala ngqo nge-ejenti yokuzincibilikisa ngaphandle kokukhuliswa.
Umsebenzi wokuhlola ulula: umjikelezo wokuqala womsebenzi wokukhulisa ulula, kanti ukwakhiwa komtapo wolwazi kuqedwa emahoreni ama-3, okuthuthukisa ukusebenza kahle komsebenzi.
Ukuthayipha okuphelele kanye nokumelana nezidakamizwa: ukumboza izindawo zokuthayipha kanye nokumelana nezidakamizwa ze-MTB kanye ne-NTM, okuyizinto ezibalulekile ezikhathazayo ngokwezokwelapha, ukuthayipha okunembile kanye nokutholwa kokumelana nezidakamizwa, ukusekela isofthiwe yokuhlaziya ezimele, kanye nokukhiqiza imibiko yokuhlaziya ngokuchofoza okukodwa.
Ukuhambisana: ukuhambisana komkhiqizo, ukuzivumelanisa nezinkundla ezijwayelekile ze-ILM kanye ne-MGI/ONT.
Imininingwane yomkhiqizo
| Ikhodi yomkhiqizo | igama lomkhiqizo | Ipulatifomu yokuthola | imininingwane |
| I-HWTS-NGS-T001 | Ikhithi yokuthola ukuthayipha kwe-Mycobacterium kanye nokumelana nemithi (indlela yokukhulisa i-multiplex) | ONT, Illumina, MGI, Salus pro | 16/96rxn |
Isikhathi sokuthunyelwe: Jan-23-2024