Trypanosome SL-RNA detection in blood and cerebrospinal fluid to demonstrate active gambiense human African trypanosomiasis infection
| العنوان: | Trypanosome SL-RNA detection in blood and cerebrospinal fluid to demonstrate active gambiense human African trypanosomiasis infection |
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| المؤلفون: | Pati Pyana, Veerle Lejon, Ipos Ngay Lukusa, Nick Van Reet, Erick Mwamba Miaka, Philippe Büscher, Felix Akwaso, Dieudonné Mumba Ngoyi, Wilfried Mutombo, Lewis Kaninda, Medard Ilunga, Justin Masumu, Vincent Kobo, Sandra Rembry, Antoine Tarral, Olaf Valverde Mordt, Digas Ngolo, Sylvain Mutanda, Dieudonné Mpoyi Muamba |
| المصدر: | PLoS Neglected Tropical Diseases, Vol 15, Iss 9, p e0009739 (2021) PLoS Neglected Tropical Diseases |
| بيانات النشر: | Public Library of Science (PLoS), 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Physiology, Trypanosoma brucei gambiense, RC955-962, Nervous System, law.invention, Cerebrospinal fluid, Medical Conditions, law, Zoonoses, Arctic medicine. Tropical medicine, Medicine and Health Sciences, African trypanosomiasis, Polymerase chain reaction, Cerebrospinal Fluid, Protozoans, Eukaryota, Body Fluids, Blood, Infectious Diseases, Democratic Republic of the Congo, RNA extraction, Lymph, Anatomy, Public aspects of medicine, RA1-1270, RNA, Protozoan, Research Article, Neglected Tropical Diseases, Trypanosoma, African Trypanosomiasis, Extraction techniques, Trypanosomiasis, medicine, Parasitic Diseases, Humans, Protozoan Infections, business.industry, Public Health, Environmental and Occupational Health, Organisms, RNA, Biology and Life Sciences, medicine.disease, Tropical Diseases, Virology, Reverse transcriptase, Parasitic Protozoans, Research and analysis methods, Trypanosomiasis, African, Parasitology, business |
| الوصف: | Background Spliced Leader (SL) trypanosome RNA is detectable only in the presence of live trypanosomes, is abundant and the Trypanozoon subgenus has a unique sequence. As previously shown in blood from Guinean human African trypanosomiasis (HAT) patients, SL-RNA is an accurate target for diagnosis. Detection of SL-RNA in the cerebrospinal fluid (CSF) has never been attempted. In a large group of Congolese gambiense HAT patients, the present study aims i) to confirm the sensitivity of SL-RNA detection in the blood and; ii) to assess the diagnostic performance of SL-RNA compared to trypanosome detection in CSF. Methodology/Principal findings Blood and CSF from 97 confirmed gambiense HAT patients from the Democratic Republic of Congo were collected using PAXgene blood RNA Tubes. Before RNA extraction, specimens were supplemented with internal extraction control RNA to monitor the extraction, which was performed with a PAXgene Blood RNA Kit. SL-RNA qPCR was carried out with and without reverse transcriptase to monitor DNA contamination. In blood, 92/97 (94.8%) HAT patients tested SL-RNA positive, which was significantly more than combined trypanosome detection in lymph and blood (78/97 positive, 80.4%, p = 0.001). Of 96 CSF RNA specimens, 65 (67.7%) were SL-RNA positive, but there was no significant difference between sensitivity of SL-RNA and trypanosome detection in CSF. The contribution of DNA to the Cq values was negligible. In CSF with normal cell counts, a fraction of SL-RNA might have been lost during extraction as indicated by higher internal extraction control Cq values. Conclusions/Significance Detection of SL-RNA in blood and CSF allows sensitive demonstration of active gambiense HAT infection, even if trypanosomes remain undetectable in blood or lymph. As this condition often occurs in treatment failures, SL-RNA detection in blood and CSF for early detection of relapses after treatment deserves further investigation. Trial registration This study was an integral part of the diagnostic trial "New Diagnostic Tools for Elimination of Sleeping Sickness and Clinical Trials: Early tests of Cure" (DiTECT-HAT-WP4, ClinicalTrials.gov Identifier: NCT03112655). Author summary Human African trypanosomiasis is a parasitic infection occurring in sub-Saharan Africa, which is fatal if left untreated. Diagnosis relies on demonstration of trypanosomes, which may occur at such low concentrations that they remain microscopically undetectable. Nucleic acid detection offers an alternative, in particular RNA, which is unstable and a better marker for live organisms than DNA. Trypanosomal SL-RNA detection in blood by reverse transcriptase quantitative PCR has hitherto only been tested twice. Although in cerebrospinal fluid, trypanosome presence indicates brain infection, SL-RNA detection has never been attempted. We evaluated sensitivity of SL-RNA detection in blood and cerebrospinal fluid. For each specimen, 2 controls were included: presence of genomic DNA contamination and efficacy of RNA extraction. Sensitivity of SL-RNA detection in blood was higher than of combined blood and lymph microscopy. In cerebrospinal fluid, SL-RNA and trypanosome detection had similar sensitivity. In a few specimens, traces of DNA were amplified. In some cerebrospinal fluids, some RNA was lost during extraction. Performing both internal controls is crucial, to ensure that negative SL-RNA cerebrospinal fluid findings are not due to a failed extraction and, in particular when testing treated patients, to differentiate live parasite RNA from reminiscent DNA. |
| اللغة: | English |
| تدمد: | 1935-2735 1935-2727 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3ff655d1c281e963925e6f651fc42940 https://doaj.org/article/fdf427a85f3c4ba9b61b8c85df1439bd |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....3ff655d1c281e963925e6f651fc42940 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19352735 19352727 |
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