Young children cannot easily produce sputum for diagnosis of pulmonary tuberculosis. Alternatively, Mycobacterium tuberculosis complex (MTB) bacilli can be detected in stool by using the Xpert MTB/RIF (Ultra) assay (Xpert). Published stool processing methods contain somewhat complex procedures and additional supplies. The aim of this study was to develop a simple one step (SOS) stool processing method, based on gravity sedimentation only, similar to sputum Xpert testing for the detection of MTB in stool. We first assessed if the SOS stool method could provide valid Xpert results without the need of bead-beating, dilution and filtration steps. We concluded that this was the case and, subsequently, validated the SOS stool method by testing spiked stool samples. By using the SOS stool method, of the 29 spiked samples, 27 gave valid Xpert results and MTB was recovered from all 27. The proof of principle of the SOS stool method was demonstrated in a routine setting in Addis Ababa, Ethiopia. Nine of 123 children with presumptive TB were MTB positive on nasogastric aspirate (NGA), and seven (77.8%) of these also had an MTB positive Xpert result on stool. Additionally, MTB was detected in the stool but not on the NGA of two children. The SOS stool processing method makes use of the standard Xpert assay kit, without the need for additional supplies or equipment. The method can potentially be rolled out to any Xpert site, bringing a bacteriologically confirmed diagnosis of TB in children closer to the point of care.
Tuberculosis, human immunodeficiency virus and hepatitis B and C infections are serious public health problems in Ethiopia. This study was conducted to assess the prevalence of triple viral infections and associated factors among tuberculosis patients who were on anti-tuberculosis treatment in the year 2011/12 in West Arsi Zone, Ethiopia. A cross sectional study design was conducted among 374 TB patients who were on first line anti-TB treatment. Simple random sampling technique was employed to draw the study subjects until the calculated sample size was achieved. Blood samples were collected to determine the HIV serostatus by rapid HIV testing kits; whereas the hepatitis B and C sero-status was determined by instant one step HBsAg and Flavicheck-HCV WB kits, respectively. Socio-demographic and risk factors information was collected and analysed using univariate and multivariate logistic regression. Injection drug users were excluded from this study. From a total of 374 tuberculosis patients enrolled, TB-HIV co-infection was found to be 56 (14.97%) among which 5 (8.92%) hepatitis B and 4 (7.14%) were hepatitis C triple infected. The overall TB-HIV-Hepatitis triple infection prevalence was 2.4% (9/374). Though the current level of triple viral infections among TB patients seems to be low, an integrated program for treatment, prevention and control strategy is crucial to be considered in TB patient management.
Background The Xpert MTB/RIF Ultra (Xpert-Ultra) assay provides timely results with good sensitivity and acceptable specificity with stool specimens in children for bacteriological confirmation of tuberculosis (TB). This study aims to optimize the Simple One-Step (SOS) stool processing method for testing stool specimens using the Xpert-Ultra in children and adults in selected health facilities in Addis Ababa, Ethiopia. The study is designed to assess the robustness of the SOS stool method, to help fine-tune the practical aspects of performing the test and to provide insights in stool storage conditions and sampling strategies before the method can be implemented and scaled in routine settings in Ethiopia as well as globally. Methods and design The project “painless optimized diagnosis of TB in Ethiopian children” (PODTEC) will be a cross sectional study where three key experiments will be carried out focusing on 1) sampling strategy to investigate if the Xpert-Ultra M. tuberculosis (MTB) -positivity rate depends on stool consistency, and if sensitivity can be increased by taking more than one stool specimen from the same participant, or doing multiple tests from the same stool specimen, 2) storage conditions to determine how long and at what temperature stool can be stored without losing sensitivity, and 3) optimization of sensitivity and robustness of the SOS stool processing method by varying stool processing steps, stool volume, and storage time and conditions of the stool-sample reagent mixture. Stool specimens will be collected from participants (children and adults) who are either sputum or naso-gastric aspiration (NGA) and/or stool Xpert-Ultra MTB positive depending on the experiment. Stool specimens from these participants, recruited from 22 sites for an ongoing related study, will be utilized for the PODTEC experiments. The sample size is estimated to be 50 participants. We will use EpiData for data entry and Stata for data analysis purposes. The main analyses will include computing the loss or gain in the Xpert-Ultra MTB positivity rate and rates of non-determinate Xpert-Ultra test results per experiment compared to the Xpert-Ultra MTB result of stool processed according to the published standard operating procedures for SOS stool processing. The differences in the MTB positivity rate by regarding testing more than one sample per child, and using different storage, and processing conditions, will be also compared to the baseline (on-site) Xpert-Ultra result.
Background Xpert MTB/RIF Ultra (Xpert-Ultra) provides timely results with good sensitivity and acceptable specificity with stool samples in children for bacteriological confirmation of tuberculosis (TB). This study aims to optimize the Simple One-Step (SOS) stool processing method for testing stool samples using the Xpert-Ultra in children and adults in selected health facilities in Addis Ababa, Ethiopia. The study is designed to assess the robustness of the SOS stool method, to help fine-tune the practical aspects of performing the test and to provide insights in stool storage conditions and sampling strategies before the method can be implemented and scaled in routine settings in Ethiopia as well as globally. Methods and design The project “painless optimized diagnosis of TB in Ethiopian children” (PODTEC) will be a cross sectional study where three key experiments will be carried out focusing on 1) sampling strategy to investigate if the Xpert-Ultra M.tuberculosis (MTB) -positivity rate depends on stool consistency, and if sensitivity can be increased by taking more than one stool sample from the same participant, or doing multiple tests from the same stool sample, 2) storage conditions to determine how long and at what temperature stool can be stored without losing sensitivity, and 3) optimization of sensitivity and robustness of the SOS stool processing method by varying sample processing steps, stool volume, and sampling from the stool-sample reagent mixture. Stool samples will be collected from participants (children and adults) who are either sputum or naso-gastric aspiration (NGA) and/or stool Xpert-Ultra MTB positive depending on the experiment. Stool samples from these participants, recruited from 22 sites for an ongoing related study, will be utilized for the PODTEC experiments. The sample size is estimated will be 50 participants. We will use EpiData for data entry and Stata for data analysis purposes. The main analyses will include computing the loss or gain in the Xpert-Ultra MTB positivity rate, and rates of unsuccessful test results. The differences in the positivity rate regarding testing more than one sample per child, different storage, and processing conditions, will be compared to the baseline (on-site) Xpert-Ultra result. Ethics and dissemination The protocol was reviewed and approved by the Ethical Review Board of the Ethiopian Public Health Institute (EPHI-IRB) (Protocol no EPHI-IRB-234-2020). The study results will be shared with the national TB program and stakeholders to the benefit of further roll out of the test in a routine Ethiopian setting. The results will also be disseminated in peer-reviewed scientific journals.
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