Reflex Xpert MTB/XDR Testing of Residual Rifampicin-Resistant Specimens: A Clinical Laboratory-Based Diagnostic Accuracy and Feasibility Study in South Africa

Author:

Centner C M12,Munir R3,Tagliani E4,Rieß F5,Brown P1,Hayes C6,Dolby T7,Zemanay W1,Cirillo D M4,David A3,Schumacher S G8,Denkinger C M89ORCID,Ruhwald M8,Leukes V N8,Nicol M P10,Van der Walt I6,Kisten G7,Gumede M7,Mace A8,Brink A1211,Stevens W312,Scott L312,Penn-Nicholson A8ORCID,Cox H111ORCID, ,Leukes Vinzeigh,Penn-Nicholson Adam,Ruhwald Morten,Erkosar Berra,Schumacher Samuel G,Singh Sunita,Kivuma Bernard,Nuru Muhuminu,Mlenge Judith,Shija Neema,Bulime Deogratias,Mnzava Dorcas,Sabuni Petro,Temba Hosiana,Siru Jamali,Hella Jerry,Msafiri Jonathan,Weisser Maja,Mbaruku Mohamed,Sasamalo Mohamed,Leonard Alice,Malango Ambilikile,Alexander Annastazia,Komakoma Faith,Msigala Gloria,Johaness Kasmir,Mhalu Grace,Hamis Mwajabu,Mlay Priscilla,Ndege Robert,Barasa Sera,Masoud Swalehe,Byakuzana Theonestina,Lwilla Anange,Kayombo Benedict,Mangu Chacha,Manyama Christina,Mbunda Theodora,Siyame Elimina,Sabi Issa,Mwaipopo Last,Ntinginya Nyanda Elias,Edom Raphael,Olomi Willyhelmina,Elisio Delio,Nguenha Dinis,Mambuque Edson,Cossa Joaquim,Cossa Marta,Gomes Neide,Manjate Patricia,Munguambe Shilzia,Acacio Sozinho,Saavedra Belen,Chiconela Helio,Ribeiro Katia,Machiana António,Meggi Bindiya,Junior Candido Azize,Madeira Carla,Khosa Celso,Bila Claudio,Floripes Denise,Malamule Diosdélio,Viegas Sofia,Saavedra Belén,Amroune Carole,Ehrlich Joanna,de la Torre Pérez Laura,Sanz Sergi,Garcia-Basteiro Albero,Riess Friedrich,Mutuku Sarah,Appalarowthu Tejaswi,Larson Leyla,Kranzer Katharina,Hoelscher Michael,Heinrich Norbert,del Mar Castro Noriega Maria,Denkinger Claudia M,Arif Saima,Cirillo Daniela Maria,Tagliani Elisa,Di Marco Federico,Batignani Virginia,Malhotra Akash,Dowdy David,Schacht Claudia,Buech Julia,Stöhr Caroline,Loembé Marguerite Massinga,Ondoa Pascale,Ndlovu Nqobile,Brown Fumbani,Ghebrekristos Yonas,Hayes Cindy,Van der Walt Ilse,Abrahams Shareef,Marokane Puleng,Radebe Mbuti,Martinson Neil,David Anura,Scott Lesley,Ngwenya Lucky,Da Silva Pedro,Solomon Reyhana,Stevens Wendy,Abongomera Charles,Reither Klaus,Stieger Leon,Brink Adrian,Centner Chad M,Cox Helen,van Heerden Judi,Nicol Mark P,Hapeela Nchimunya,Brown Parveen,Solomon Reyhana,Zemanay Widaad,Dolby Tania

Affiliation:

1. Division of Medical Microbiology, Department of Pathology, University of Cape Town , Cape Town , South Africa

2. National Health Laboratory Service, Groote Schuur Hospital , Cape Town , South Africa

3. Wits Diagnostic Innovation Hub, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand , Johannesburg , South Africa

4. Emerging Bacterial Pathogens Unit, IRCCS Ospedale San Raffaele , Milan , Italy

5. Division of Infectious Diseases and Tropical Medicine, LMU University Hospital, LMU Munich , Munich , Germany

6. National Health Laboratory Service , Gqeberha , South Africa

7. National Health Laboratory Service , Cape Town , South Africa

8. Tuberculosis Programme, FIND , Geneva , Switzerland

9. Division of Infectious Disease and Tropical Medicine, Center for Infectious Diseases, Heidelberg University Hospital, and German Center for Infection Research (DZIF), Partner Site Heidelberg , Heidelberg , Germany

10. Marshall Centre for Infectious Diseases Research and Training, School of Biomedical Sciences, University of Western Australia , Perth, Western Australia , Australia

11. Welcome Centre for Infectious Disease Research in Africa and Institute of Infectious Disease and Molecular Medicine, University of Cape Town , Cape Town , South Africa

12. National Priority Program, National Health Laboratory Service , Johannesburg , South Africa

Abstract

Abstract Background The World Health Organization–approved Xpert MTB/XDR test detects Mycobacterium tuberculosis and resistance to isoniazid, fluoroquinolones, ethionamide, and injectable drugs directly in specimens. This pragmatic, laboratory-based study assessed the diagnostic accuracy and feasibility of a reflex testing approach, where Xpert MTB/XDR was performed on residual specimens previously processed for Xpert MTB/RIF Ultra. Methods Routine respiratory specimens, processed for Xpert MTB/RIF Ultra, were stored in sample reagent buffer at 2°C–8°C. If rifampicin resistant, the residual specimen was assessed for adequate volume (≥2 mL) and tested with Xpert MTB/XDR, with storage time recorded. A second specimen was used for routine and reference standard testing (culture and sequencing). Results Specimens (99% sputum) from 763 participants submitted to 2 large routine laboratories were included. Xpert MTB/XDR yielded valid resistance detection results in 639 (84%), compared with 507 (66%) for routine testing (difference [95% CI], 18% [13%–22%]). The median turnaround time for results was 23 hours for Xpert MTB/XDR and 15 days for routine testing. While 748 specimens (98%) were ≥2 mL, only 102 (13%) were stored for ≤4 hours. By the reference standard, 284 of 394 (72%) were isoniazid resistant, and 57 of 380 (15%) were fluroquinolone resistant. The sensitivities of Xpert MTB/XDR were 94% (95% CI, 91%–97%) for isoniazid and 91% (81%–97%) for fluoroquinolone resistance detection. The specificities were 98% (94%–100%) and 100% (98%–100%), respectively. Conclusions Xpert MTB/XDR performed favorably compared with the reference, and the reflex testing approach increased results availability over routine testing, while dramatically decreasing turnaround time from weeks to hours. Laboratory workflow precluded testing within the manufacturer-recommended 4-hour storage time, but longer storage did not appear detrimental.

Funder

European Union

Publisher

Oxford University Press (OUP)

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