CRISPR detection of circulating cell-free Mycobacterium tuberculosis DNA in adults and children, including children with HIV: a molecular diagnostics study

被引:67
作者
Huang, Zhen [1 ,2 ,3 ]
LaCourse, Sylvia M. [4 ,5 ]
Kay, Alexander W. [7 ,8 ]
Stern, Joshua [5 ]
Escudero, Jaclyn N. [5 ]
Youngquist, Brady M. [1 ,2 ]
Zheng, Wenshu [1 ,2 ]
Vambe, Debrah [9 ]
Dlamini, Muyalo [10 ]
Mtetwa, Godwin [7 ,8 ]
Cranmer, Lisa M. [11 ,12 ,13 ]
Njuguna, Irene [5 ,14 ]
Wamalwa, Dalton C. [5 ,15 ]
Maleche-Obimbo, Elizabeth [5 ,15 ]
Catanzaro, Donald G. [16 ]
Lyon, Christopher J. [1 ,2 ]
John-Stewart, Grace [4 ,5 ,6 ]
DiNardo, Andrew [7 ]
Mandalakas, Anna M. [7 ]
Ning, Bo [1 ,2 ]
Hu, Tony Y. [1 ,2 ]
机构
[1] Tulane Univ, Sch Med, Ctr Cellular & Mol Diagnost, New Orleans, LA 70112 USA
[2] Tulane Univ, Sch Med, Dept Biochem & Mol Biol, New Orleans, LA 70112 USA
[3] Nanchang Univ, State Key Lab Food Sci & Technol, Nanchang, Jiangxi, Peoples R China
[4] Univ Washington, Dept Med, Div Allergy & Infect Dis, Seattle, WA 98195 USA
[5] Univ Washington, Dept Med, Dept Global Hlth, Seattle, WA 98195 USA
[6] Univ Washington, Dept Med, Dept Pediat, Seattle, WA 98195 USA
[7] Baylor Coll Med, Dept Pediat, Global TB Program, Houston, TX 77030 USA
[8] Baylor Childrens Fdn Eswatini, Mbabane, Eswatini
[9] Minist Hlth, Eswatini Natl TB Control Programme, Manzini, Eswatini
[10] Minist Hlth, Eswatini Hlth Lab Serv, NationalTB Reference Lab, Mbabane, Eswatini
[11] Emory Univ, Dept Pediat, Div Infect Dis, Atlanta, GA 30322 USA
[12] Childrens Healthcare Atlanta, Atlanta, GA USA
[13] Emory Rollins Sch Publ Hlth, Dept Epidemiol, Atlanta, GA USA
[14] Kenyatta Natl Hosp, Res & Programmes, Nairobi, Kenya
[15] Univ Nairobi, Dept Paediat & Child Hlth, Nairobi, Kenya
[16] Univ Arkansas, Dept Biol Sci, Fayetteville, AR 72701 USA
关键词
XPERT MTB/RIF ASSAY; PULMONARY TUBERCULOSIS; PREVALENCE; URINE;
D O I
10.1016/S2666-5247(22)00087-8
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
Background Tuberculosis remains a leading cause of global mortality, especially for adults and children living with HIV (CLHIV) underdiagnosed by sputum-based assays. Non-sputum-based assays are needed to improve tuberculosis diagnosis and tuberculosis treatment monitoring. Our aim in this study was to determine whether ultrasensitive detection of Mycobacterium tuberculosis cell-free DNA (Mtb-cfDNA) in blood can diagnose tuberculosis and evaluate tuberculosis treatment responses. Methods In this molecular diagnostics study we analysed archived serum from two patient populations evaluated for tuberculosis in Eswatini and Kenya to detect Mtb-cfDNA, analysing serum from all individuals who had both sufficient serum volumes and clear diagnostic results. An optimised CRISPR-mediated tuberculosis (CRISPR-TB) assay was used to detect Mtb-cfDNA in serum at enrolment from adults and children with presumptive tuberculosis and their asymptomatic household contacts, and at enrolment and during tuberculosis treatment from a cohort of symptomatic CLHIV at high risk for tuberculosis, who provided longitudinal serum at enrolment and during tuberculosis treatment. Findings CRISPR-TB identified microbiologically and clinically confirmed tuberculosis cases in the predominantly HIV-negative Eswatini adult cohort with 96% sensitivity (27 [96%] of 28, 95% CI 80-100) and 94% specificity (16 [94%] of 17, 71-100), and with 83% sensitivity (5 [83%] of 6, 36-100) and 95% specificity (21 [95%] of 22, 77-100) in the paediatric cohort, including all six cases of extrapulmonary tuberculosis. In the Kenyan CLHIV cohort, CRISPR-TB detected all (13 [100%] of 13, 75-100) confirmed tuberculosis cases and 85% (39 [85%] of 46, 71-94) of unconfirmed tuberculosis cases diagnosed by non-microbiological clinical findings. CLHIV who were CRISPR-TB positive at enrolment had a 2 center dot 4-times higher risk of mortality by 6 months after enrolment. Mtb-cfDNA signal decreased after tuberculosis treatment initiation, with near or complete Mtb-cfDNA clearance by 6 months after tuberculosis treatment initiation. Interpretation CRISPR-mediated detection of circulating Mtb-cfDNA shows promise to increase the identification of paediatric tuberculosis and HIV-associated tuberculosis, and potential for early diagnosis and rapid monitoring of tuberculosis treatment responses.
引用
收藏
页码:482 / 492
页数:11
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