Isixhobo Esitsha Sokuxilongwa Kwesifo Sephepha kunye Nokuchongwa Kokuchasana Neziyobisi: Ulandelelwano Olujoliswe Kwisizukulwana Esitsha (tNGS) Oludityaniswe Nokufunda Koomatshini Ukuxilongwa Kwesifo Sephepha
Ingxelo yoncwadi: I-CCa: imodeli yokuxilonga esekelwe kwi-tNGS kunye nokufunda koomatshini, efanelekileyo kubantu abanesifo sephepha esincinci sebhaktiriya kunye ne-tuberculous meningitis.
Isihloko sethisisi: Ulandelelwano lwesizukulwana esilandelayo olujoliswe kwisifo sofuba kunye nokufunda koomatshini: icebo lokuxilonga elinobuzaza kakhulu kwiityhubhu zemiphunga ezibuthathaka kunye ne-meningitis ejikelezayo.
Ngamaxesha athile: 《Clinica Chimica Acta》
UKUBA: 6.5
Umhla wokupapashwa: uJanuwari 2024
Idibene neYunivesithi yaseTshayina yeSayensi kunye neBeijing Chest Hospital yeCapital Medical University, iMacro & Micro-Test iseke imodeli yokuxilonga isifo sephepha esekelwe kwisizukulwana esitsha setekhnoloji yokulandelelana okujoliswe kuko (tNGS) kunye nendlela yokufunda komatshini, ebonelele ngokuqonda okuphezulu kakhulu kwesifo sephepha esineebhaktheriya ezimbalwa kunye ne-tuberculous meningitis, ibonelele ngendlela entsha yokuxilonga isifo sephepha esibangelwa kukunganyamezelani kwiintlobo ezimbini zesifo sephepha, kwaye yanceda ekuxilongweni ngokuchanekileyo, ekufumaneni ukunganyangeki ngamayeza kunye nonyango lwesifo sephepha. Kwangaxeshanye, kufunyaniswe ukuba i-plasma cfDNA yesigulana ingasetyenziswa njengohlobo olufanelekileyo lwesampulu yokuvavanya isifo sephepha ekuxilongweni kweTBM.
Kolu phononongo, iisampulu zeplasma ezingama-227 kunye neesampulu zolwelo lwe-cerebrospinal zasetyenziswa ukuseka amaqela amabini eklinikhi, apho iisampulu zeqela lokuxilongwa kwelabhoratri zasetyenziswa ukuseka imodeli yokufunda yomatshini yokuxilongwa kwesifo sephepha, kwaye iisampulu zeqela lokuxilongwa kweklinikhi zasetyenziswa ukuqinisekisa imodeli yokuxilonga esekwe. Zonke iisampulu zaqala zajoliswa yidama le-capture probe elijoliswe ngokukodwa leMycobacterium tuberculosis. Emva koko, ngokusekelwe kwidatha yokulandelelana kwe-TB-tNGS, imodeli yomthi wesigqibo isetyenziselwa ukwenza ukuqinisekiswa okuphindwe kahlanu kwiiseti zoqeqesho kunye nokuqinisekiswa komgca wokuxilonga welabhoratri, kwaye kufunyanwa imida yokuxilonga yesampulu zeplasma kunye neesampulu zolwelo lwe-cerebrospinal. Umda ofunyenweyo uziswa kwiiseti ezimbini zovavanyo lomgca wokuxilonga weklinikhi ukuze kufunyanwe, kwaye ukusebenza komfundi kuhlolwe yi-ROC curve. Ekugqibeleni, imodeli yokuxilonga yesifo sephepha yafunyanwa.
Umzobo 1 umzobo wesicwangciso soyilo lophando
Iziphumo: Ngokwemida ethile yesampuli ye-CSF DNA (AUC = 0.974) kunye nesampuli ye-plasma cfDNA (AUC = 0.908) echazwe kolu phononongo, phakathi kweesampuli ezingama-227, uvakalelo lwesampuli ye-CSF yayiyi-97.01%, uvakalelo yayiyi-95.65%, kwaye uvakalelo kunye novakalelo lwesampuli ye-plasma yayiyi-82.61% kunye ne-86.36%. Kuhlalutyo lweesampuli ezingama-44 ezidibeneyo ze-plasma cfDNA kunye ne-cerebrospinal fluid DNA ezivela kwizigulane ze-TBM, icebo lokuxilonga lolu phononongo linokuhambelana okuphezulu kwe-90.91% (40/44) kwi-plasma cfDNA kunye ne-cerebrospinal fluid DNA, kwaye uvakalelo yi-95.45% (42/44). Kubantwana abanesifo sephepha semiphunga, icebo lokuxilonga lolu phononongo livakalelwa ngakumbi kwiisampuli ze-plasma kuneziphumo zokufumanisa ze-Xpert zeesampuli zejusi yesisu ezivela kwizigulane ezifanayo (28.57% VS 15.38%).
Umzobo 2 Uhlalutyo lokusebenza kwemodeli yokuchongwa kwesifo sephepha kwiisampulu zabemi
Umzobo 3 Iziphumo zokuxilonga zeesampuli ezibhangqiweyo
Isiphelo: Indlela yokuxilonga isifo sephepha ebuthathaka kakhulu yasekwa kolu phononongo, enokubonelela ngesixhobo sokuxilonga esinobukrelekrele bokubona obuphezulu kwizigulana zonyango ezine-oligobacillary tuberculosis (negative culture). Ukufumanisa isifo sephepha esibuthathaka kakhulu ngokusekelwe kwi-plasma cfDNA kunokuba luhlobo olufanelekileyo lwesampulu yokuxilonga isifo sephepha esisebenzayo kunye ne-tuberculous meningitis (iisampulu ze-plasma kulula ukuziqokelela kune-cerebrospinal fluid kwizigulana ezirhanelwa ukuba zinesifo sephepha sobuchopho).
Ikhonkco lokuqala: https://www.sciencedirect.com/science/article/pii/s0009898123004990? via%3Dihub
Intshayelelo emfutshane yeemveliso zokufumanisa uthotho lwesifo sephepha seMacro kunye neMicro-Test
Ngenxa yemeko enzima yesampulu yezigulana zesifo sofuba kunye neemfuno ezahlukeneyo, iMacro & Micro-Test ibonelela ngeseti epheleleyo yezisombululo ze-NGS zokukhupha ulwelo kwiisampuli zesikhohlela, ukwakhiwa kunye nokulandelelana kwethala leencwadi leQualcomm, kunye nohlalutyo lwedatha. Ezi mveliso ziquka ukuxilongwa ngokukhawuleza kwezigulana zesifo sofuba, ukufunyanwa kwesifo sofuba esinganyangekiyo ngamayeza, uhlobo lwesifo sofuba se-mycobacterium kunye ne-NTM, ukuxilongwa kwesifo sofuba esingenabhaktheriya kunye nabantu abanesi sifo sofuba, njl.
Izixhobo zokufumanisa isifo sephepha kunye ne-mycobacteria:
| Inombolo yento | Igama lemveliso | umxholo wovavanyo lwemveliso | uhlobo lwesampulu | imodeli efanelekileyo |
| I-HWTS-3012 | Iarhente yokukhupha isampuli | Isetyenziswa kunyango lokunyibilikisa iisampuli zesikhohlela, ifumene inombolo yerekhodi yeklasi yokuqala, iSutong Machinery Equipment 20230047. | isikhwehlela | |
| I-HWTS-NGS-P00021 | Isixhobo sokufumanisa ubungakanani beQualcomm sesifo sephepha esinovakalelo olukhulu (indlela yokubamba iprobe) | Ukufunyanwa kwe-hypersensitivity engangenisi (i-biopsy yolwelo) kwi-TB yemiphunga kunye nee-nodules zobuchopho ezingenayo intsholongwane; Iisampulu zabantu ekurhanelwa ukuba bosulelwe sisifo sephepha okanye i-mycobacteria engeyiyo i-TB zihlalutywe nge-metagenomics enzulu, kwaye ulwazi lokufumanisa ukuba i-mycobacteria yesifo sephepha okanye engeyiyo i-TB zisulelwe na kwaye ulwazi oluphambili lokumelana namayeza kwi-mycobacterium tuberculosis lubonelelwe. | Igazi elingaphandle, ulwelo lokuhlamba i-alveolar, i-hydrothorax kunye ne-ascites, isampuli yokubhoboza okugxile, ulwelo lwe-cerebrospinal. | Isizukulwana sesibini |
| I-HWTS-NGS-T001 | Ikhithi yokuchonga ukuchasana neziyobisi zeMycobacterium (indlela yokulandelelanisa i-multiplex amplification) | Uvavanyo lokuchwetheza lweMycobacterium, kuquka iMTBC kunye ne-187 NTM；Ukufunyanwa kweMycobacterium tuberculosis ukumelana namayeza kuquka amayeza ali-13 kunye neendawo ezili-16 eziphambili zotshintsho lwezakhi zofuzo ukumelana namayeza. | I-sputum, ulwelo lokuhlamba i-alveolar, i-hydrothorax kunye ne-ascites, isampuli yokubhoboza okugxile, ulwelo lwe-cerebrospinal. | Iqonga lesibini/lesithathu lesizukulwana esiphindwe kabini |
Izinto ezibalaseleyo: Ikhithi yokuchonga kunye nokuchonga ukumelana namayeza ye-HWTS-NGS-T001 Mycobacterium (indlela yokukhulisa i-multiplex)
Intshayelelo yemveliso
Le mveliso isekelwe kumayeza aphambili okuqala nowesibini achazwe kwizikhokelo zonyango lwe-WHO TB, ii-macrolides kunye nee-aminoglycosides ezisetyenziswa kakhulu kwizikhokelo zonyango lwe-NTM, kwaye iindawo zokuxhathisa amayeza zigubungela lonke iqela elinye leendawo ezinxulumene nokuxhathisa amayeza kwikhathalogu ye-WHO Mycobacterium tuberculosis complex mutation, kunye nezinye ii-genes ezixeliweyo kunye neendawo zokutshintsha amayeza ngokophando kunye nezibalo zeencwadi ezibalaseleyo ekhaya nakwamanye amazwe.
Ukuchonga uhlobo lwe-NTM kusekelwe kwiintlobo ze-NTM ezishwankathelwe kwizikhokelo ze-NTM ezipapashwe yiChinese Journal of Tuberculosis and Respiratory Diseases kunye nemvumelwano yeengcali. Iiprimer zokubhala eziyilwe ngobuchule zinokwandisa, zilandelelanise kwaye zichaze iintlobo ze-NTM ezingaphezu kwe-190.
Ngokusebenzisa iteknoloji yokukhulisa i-multiplex PCR ekujoliswe kuyo, ii-genotyping genes kunye nee-genes ezichasene namayeza zeMycobacterium zandiswa yi-multiplex PCR, kwaye indibaniselwano ye-amplicon yee-genes ekujoliswe kuzo eza kufunyanwa yafunyanwa. Iimveliso eziphuculweyo zinokwakhiwa kwiilayibrari zokulandelelana kwesizukulwana sesibini okanye sesithathu, kwaye zonke iiplatifomu zokulandelelana kwesizukulwana sesibini nesesithathu zinokujongwa ngokulandelelana okunzulu ukuze kufunyanwe ulwazi lokulandelelana kwee-genes ekujoliswe kuzo. Ngokuthelekisa ne-mutations ezaziwayo ezikwi-database yesalathiso esakhelwe ngaphakathi (kubandakanya ikhathalogu ye-WHO Mycobacterium tuberculosis complex mutation kunye nobudlelwane bayo nokunganyamezelani namayeza), i-mutations enxulumene nokunganyamezelani namayeza okanye ukuthambekela kwamayeza okulwa nesifo sephepha yafunyanwa. Idityaniswe nesisombululo sonyango lwesampuli ye-sputum esizivulekeleyo seMacro & Micro-Test, ingxaki yokusebenza kakuhle kwe-nucleic acid amplification yeesampuli ze-sputum zeklinikhi (eziphindwe kalishumi kuneendlela zemveli) yasonjululwa, ukuze ukufunyanwa kokulandelelana kokunganyamezelani namayeza kusetyenziswe ngokuthe ngqo kwiisampuli ze-sputum zeklinikhi.
Uluhlu lokuchonga imveliso
34iijini ezinxulumene nokungaxhathisi amayeza18amayeza okulwa nesifo sephepha kunye6Kufunyenwe amayeza e-NTM, agubungela297iindawo zokuxhathisa amayeza; Iintlobo ezilishumi zeMycobacterium tuberculosis kunye nezinye ezingaphezulu190iintlobo ze-NTM zifunyenwe.
Itheyibhile 1: Ulwazi malunga namayeza aneminyaka eli-18+6 +190+NTM
Inzuzo yemveliso
Ukulungelelana okuqinileyo kwezonyango: iisampulu zesikhohlela zinokufunyaniswa ngokuthe ngqo nge-arhente yokuzinyibilikisa ngaphandle kokukhuliswa.
Umsebenzi wovavanyo ulula: umjikelo wokuqala womsebenzi wokwandisa ulula, kwaye ukwakhiwa kwethala leencwadi kugqitywa kwiiyure ezi-3, nto leyo ephucula ukusebenza kakuhle komsebenzi.
Ukubhala okubanzi kunye nokuxhathisa iziyobisi: ukugubungela iindawo zokubhala kunye nokuxhathisa iziyobisi ze-MTB kunye ne-NTM, ezizezona zinto zibalulekileyo kwiinkxalabo zonyango, ukuchonga ngokuchanekileyo ukuchwetheza kunye nokuxhathisa iziyobisi, ukuxhasa isoftware yohlalutyo oluzimeleyo, kunye nokuvelisa iingxelo zohlalutyo ngokucofa kube kanye.
Ukuhambelana: ukuhambelana kwemveliso, ukuziqhelanisa namaqonga aqhelekileyo e-ILM kunye ne-MGI/ONT.
Iinkcukacha zemveliso
| Ikhowudi yemveliso | Igama lemveliso | Iqonga lokubona | iinkcukacha |
| I-HWTS-NGS-T001 | Ikhithi yokuchonga ukuchasana neziyobisi zeMycobacterium (indlela yokwandisa i-multiplex) | ONT, Illumina, MGI, Salus pro | 16/96rxn |
Ixesha leposi: Jan-23-2024