Prevalence and Distribution of Multidrug-Resistant Mutations in Mycobacterium tuberculosis in Tanzania

2019 ◽  
Vol 1 (1) ◽  
pp. 15-22
Author(s):  
John C. Mgogwe ◽  
Hadija H. Semvua ◽  
Oliva Safari ◽  
Gibson E. Kapanda ◽  
Balthazar M. Nyombi ◽  
...  
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Sputum Smear ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Important Approach ◽  
Mdr Tb ◽  
Frequent Mutations ◽  
Lowenstein Jensen

Background: Molecular identification of mutations resulting in multidrug-resistant tuberculosis (MDR-TB) is an important approach for improving understanding of MDR-TB epidemiology and planning for appropriate interventions. We aimed to estimate the prevalence and distribution of mutations causing MDR-TB as well as determine the gene distribution among patients previously treated for TB. Methods: This was a cross-sectional, hospital-based study conducted from April 2017 to October 2018 at Kibong’oto Infectious Diseases Hospital (KIDH). KIDH is the national MDR-TB referral hospital. Participants were patients presumptively diagnosed with MDR-TB and referred to KIDH from district and regional hospitals across Tanzania. Sputum samples were collected and analysed using the Xpert MTB/RIF assay, direct sputum smear fluorescence microscopy, culture on Lowenstein-Jensen medium, and line probe assay using the GenoType MTBDRplus VER 2.0 system. Demographic information and mutation frequencies were reported as counts and percentages and analysed using descriptive statistics. Results: A total of 208 (69.3%) participants had rpoB gene mutations conferring resistance to only rifampicin; 92 (30.7%) had rpoB, katG, and inhA mutations conferring resistance to rifampicin and isoniazid; 78 (26%) had rpoB and katG mutations conferring resistance to rifampicin and isoniazid; and 14 (4.7%) had rpoB and inhA mutations conferring resistance to rifampicin and isoniazid. Conclusion: The mutation prevalences identified in this study indicate the most frequent mutations were the S531L mutation of the rpoB gene, the S315T1 mutation of the katG gene, and the S315T mutation in the promoter region of the inhA gene. To control the emergence and spread of MDR-TB, drug sensitivity testing must be carried for GeneXpert-confirmed TB patients prior to initiating second-line anti-TB regimens.

scholarly journals Prevalence and Distribution of Multidrug-Resistant Mutations in Mycobacterium tuberculosis in Tanzania

2019 ◽  
Vol 1 (1) ◽  
pp. 15-22
Author(s):  
John C. Mgogwe ◽  
Hadija H. Semvua ◽  
Oliva Safari ◽  
Gibson E. Kapanda ◽  
Balthazar M. Nyombi ◽  
...  
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Sputum Smear ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Important Approach ◽  
Mdr Tb ◽  
Frequent Mutations ◽  
Lowenstein Jensen

Background: Molecular identification of mutations resulting in multidrug-resistant tuberculosis (MDR-TB) is an important approach for improving understanding of MDR-TB epidemiology and planning for appropriate interventions. We aimed to estimate the prevalence and distribution of mutations causing MDR-TB as well as determine the gene distribution among patients previously treated for TB. Methods: This was a cross-sectional, hospital-based study conducted from April 2017 to October 2018 at Kibong’oto Infectious Diseases Hospital (KIDH). KIDH is the national MDR-TB referral hospital. Participants were patients presumptively diagnosed with MDR-TB and referred to KIDH from district and regional hospitals across Tanzania. Sputum samples were collected and analysed using the Xpert MTB/RIF assay, direct sputum smear fluorescence microscopy, culture on Lowenstein-Jensen medium, and line probe assay using the GenoType MTBDRplus VER 2.0 system. Demographic information and mutation frequencies were reported as counts and percentages and analysed using descriptive statistics. Results: A total of 208 (69.3%) participants had rpoB gene mutations conferring resistance to only rifampicin; 92 (30.7%) had rpoB, katG, and inhA mutations conferring resistance to rifampicin and isoniazid; 78 (26%) had rpoB and katG mutations conferring resistance to rifampicin and isoniazid; and 14 (4.7%) had rpoB and inhA mutations conferring resistance to rifampicin and isoniazid. Conclusion: The mutation prevalences identified in this study indicate the most frequent mutations were the S531L mutation of the rpoB gene, the S315T1 mutation of the katG gene, and the S315T mutation in the promoter region of the inhA gene. To control the emergence and spread of MDR-TB, drug sensitivity testing must be carried for GeneXpert-confirmed TB patients prior to initiating second-line anti-TB regimens.

Prevalence and Distribution of Multidrug-Resistant Mutations in Mycobacterium tuberculosis in Tanzania

2019 ◽  
Vol 1 (1) ◽  
pp. 15-22
Author(s):  
John C. Mgogwe ◽  
Hadija H. Semvua ◽  
Oliva Safari ◽  
Gibson E. Kapanda ◽  
Balthazar M. Nyombi ◽  
...  
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Sputum Smear ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Important Approach ◽  
Mdr Tb ◽  
Frequent Mutations ◽  
Lowenstein Jensen

Background: Molecular identification of mutations resulting in multidrug-resistant tuberculosis (MDR-TB) is an important approach for improving understanding of MDR-TB epidemiology and planning for appropriate interventions. We aimed to estimate the prevalence and distribution of mutations causing MDR-TB as well as determine the gene distribution among patients previously treated for TB. Methods: This was a cross-sectional, hospital-based study conducted from April 2017 to October 2018 at Kibong’oto Infectious Diseases Hospital (KIDH). KIDH is the national MDR-TB referral hospital. Participants were patients presumptively diagnosed with MDR-TB and referred to KIDH from district and regional hospitals across Tanzania. Sputum samples were collected and analysed using the Xpert MTB/RIF assay, direct sputum smear fluorescence microscopy, culture on Lowenstein-Jensen medium, and line probe assay using the GenoType MTBDRplus VER 2.0 system. Demographic information and mutation frequencies were reported as counts and percentages and analysed using descriptive statistics. Results: A total of 208 (69.3%) participants had rpoB gene mutations conferring resistance to only rifampicin; 92 (30.7%) had rpoB, katG, and inhA mutations conferring resistance to rifampicin and isoniazid; 78 (26%) had rpoB and katG mutations conferring resistance to rifampicin and isoniazid; and 14 (4.7%) had rpoB and inhA mutations conferring resistance to rifampicin and isoniazid. Conclusion: The mutation prevalences identified in this study indicate the most frequent mutations were the S531L mutation of the rpoB gene, the S315T1 mutation of the katG gene, and the S315T mutation in the promoter region of the inhA gene. To control the emergence and spread of MDR-TB, drug sensitivity testing must be carried for GeneXpert-confirmed TB patients prior to initiating second-line anti-TB regimens.

scholarly journals Identification of rpoB, gyrA and embB Gene Mutations in Mycobacterium Tuberculosis Isolates from Retreatment Tuberculosis Patients in Nepal

2018 ◽  
Vol 14 (2) ◽  
pp. 39-50
Author(s):  
Dhruba Kumar Khadka ◽  
Rajendra Prasad Pant ◽  
Bikash Lamichhane ◽  
Sharat Chandra Verma ◽  
R. P. Bichha ◽  
...  
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Drug Susceptibility ◽  
Cross Sectional Study ◽  
Molecular Techniques ◽  
Reference Laboratory ◽  
Drug Resistant ◽  
Cross Sectional ◽  
Mdr Tb ◽  
Embb Gene

Introduction: Tuberculosis remains one of the major public health problems in Nepal and increasing trend of multi drug resistant and extensively drug resistant tuberculosis (MDR /XDR TB) is a big challenge. Rapid diagnosis and appropriate treatment of MDR/XDR TB is crucial. Identification and comparison of MDR TB using rapid molecular techniques (for rpob, gyrA, rrs and embB gene mutations) with reference to drug susceptibility test (DST) were the main objectives of this study.Methodology: A cross sectional study was carried out in National TB Centre (NTC). Gene Xpert, proportion method and Line Probe Assay (LPA) were used for first and second line drugs susceptibility testing (FLD-DST and SLD-DST). A total of 29 mucopurulent sputum samples were freshly collected from retreatment TB patients (Female 41.4%, Male 58.6%) with median age of 40 years attending to the four MDR TB treatment centres of eastern and central Nepal (via private courier and directly to National TB Reference Laboratory (NRL) at NTC from April 2013 to October 2017.Results: Among 29 sputum samples (Female 41.4%; all smear+ve, Male 58.6%; 16 smear+ve and 1 smear-ve), Gene Xpert MTB/RIF assay detected 100% M. tuberculosis and rifampicin resistance (rpoB gene resistant) of which, 100% were culture positive by conventional Lowenstein–Jensen (LJ) method. FLDDST was performed on all culture positives of which, 96.6% showed MDR TB and 3.4% showed mono resistance to isonizid only. SLD-DST on 29 MDRTB strains by LPA showed 100%, 58.6%, 44.8% wild type for rrs, gyrA and emb B genes respectively. Mutation for gyrA and emb B genes was 41.4% and 51.2% respectively, rrs genes none. Twelve (Female 6, Male 6) MDR TB strains were identified  as pre-XDR-TB. Chi square (χ2) for trend was used to analyze Gene Xpert, smear, FLD-DST and LPA results.Conclusion: From this study, 29(100%) MDRTB were detected from retreatment TB cases by Gene Xpert and FLDDST. Almost 41.4% MDR TB strains were detected as pre-XDR TB by LPA, which were found to be higher in 15-60 years group of females and males. Samples from retreatment TB patients need to be tested by rapid molecular techniques with reference to culture and DST.SAARC Journal of Tuberculosis, Lung Diseases and HIV/AIDS, Vol. 14, No. 2, 2017, Page: 39-50

scholarly journals Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

2020 ◽  
Vol 52 (4) ◽  
Author(s):  
Linda Choerunnisa ◽  
Ida Parwati ◽  
Coriejati Rita ◽  
Anna Tjandrawati ◽  
Lidya Chaidir
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Detection Methods ◽  
Isoniazid Resistance ◽  
Resistant Tuberculosis ◽  
Position Detection ◽  
Multidrug Resistant Tuberculosis ◽  
Mdr Tb ◽  
Codon Mutation

Indonesia is one of the countries with the highest multidrug-resistant tuberculosis cases in the world. Rapid molecular test using the Xpert MTB/RIF assay is one of the detection methods for MDR-TB. Early detection of MDR-TB is crucial for early initiation of treatment. However, Xpert MTB/RIF assay only detects the rpoB gene mutations associated with Rifampicin resistance. Recently, WHO recommends the use of Pyrosequencing, a DNA sequencing method that can detect not only the rpoB gene but also katG and/or inhA gene mutations associated with Isoniazid resistance. The aims of this study were to compare the interpretation between the two methods and to determine the differences in codon mutation position detection of the rpoB gene and mutation detection of the katG and/or inhA gene. This was a cross-sectional comparative observational study on patients ≥18 years old interpreted as RR-TB patients based on Xpert MTB/RIF assay results who had not received MDR-TB drugs at Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Results showed there were 40 Rifampicin-resistant TB subjects interpreted by Xpert MTB/RIF assay while Pyrosequencing interpreted 30 MDR-TB, 9 RR-TB and one Isoniazid-resistant TB subjects in January - February 2020. The detection of rpoB gene codon mutation position between Xpert MTB/RIF assay and Pyrosequencing methods was not significantly different (p=0.389). Pyrosequencing had detected 27 katG gene mutations, 3 inhA gene mutations, one katG and inhA gene mutation. To conclude, Pyrosequencing can be used for accurate detection of Rifampicin and Isoniazid resistance in MDR-TB.

scholarly journals IDENTIFICATION GENE MUTATIONS RPOB CAUSE OF MULTIDRUG RESISTANCE TUBERCULOSIS IN HAJI ADAM MALIK HOSPITAL

2018 ◽  
Vol 11 (13) ◽  
pp. 155
Author(s):  
Adhisty Nurpermatasari ◽  
Urip Harahap ◽  
Parluhutan Siagian
Keyword(s):  
Polymerase Chain Reaction ◽  
Multidrug Resistance ◽  
General Pattern ◽  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Chain Reaction ◽  
Mdr Tb ◽  
Pcr Products ◽  
Polymerase Chain

Objective: The study aimed to investigate gene mutations in Mycobacterium tuberculosis causing resistance to rifampicin (RIF) in multidrug resistance TB patients in Haji Adam Malik Hospital and determined the suitability of the general pattern of mutations that occur in patients with multidrug-resistant TB (MDR-TB) in the hospital with global mutation pattern.Methods: This observational study was performed prospectively in 31 patients with MDR-TB who had data medical record results of GeneXpert MTB/ RIF positive for resistant to RIF period August–November 2016 in the hospital by taking sputum from patients. DNA extracted from sputum and then purified to be detected using polymerase chain reaction (PCR). PCR products were examined by electrophoresis.Results: The highest percentage of 81 bp rpoB gene mutation MTB is called as RIF -resistance determining region which was located at codon 516 (100%) and then was followed by mutation at codon 531 (96.77%) and codon 526 (90.32%), respectively, while the smallest percentage of mutation (12.90%) MTB at codon was 533.Conclusion: The rpoB gene shows a positive mutation-causing resistance to RIF.

scholarly journals Burden of drug-resistant pulmonary tuberculosis in Pakistani children: A cross-sectional study

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 344
Author(s):  
Ghulam Shabbir Laghari ◽  
Zahid Hussain ◽  
Lavina Khemani ◽  
Syed Zohaib Maroof Hussain ◽  
Uzair Yaqoob
Keyword(s):  
Drug Sensitivity ◽  
Multidrug Resistant ◽  
Sputum Smear ◽  
P Value ◽  
Drug Resistant ◽  
Duration Of Treatment ◽  
Cross Sectional ◽  
Resistant Tuberculosis ◽  
Mdr Tb ◽  
History Of

Introduction: The incidence of drug-resistant tuberculosis (TB) is rapidly increasing worldwide. Children in high TB burden countries are rapidly being reported to be affected by multidrug-resistant TB (resistant to isoniazid and rifampicin). The aim of this study is to evaluate the pattern of drug sensitivity among children suffering with TB. Methods: Known cases of pulmonary TB, with sputum smear positive even after two months of compliance to 1st line anti-tuberculous therapy were included after gaining informed consent. Specimens used for drug sensitivity analysis were either sputum or bronchoalveolar lavage. Patient age, gender, history of TB contact, and duration of treatment were also recorded. Data was entered and analyzed using SPSS v.22. Results: There were 32 male (64%) and 18 female (36%) children in the study. Their mean age was 12.84 ± 2.54 years. History of household TB contact was positive in 29 (58%) children. Among 1st line anti-tuberculous therapy, ethambutol and streptomycin were most sensitive (n=44; 88%). Rifampin was least sensitive (n=17; 34%). There were 32 (64%) children with multidrug-resistant tuberculosis (MDR-TB). A positive history of household TB contact (either resistant or non-resistant) was seen to have a statistically significant impact on incidence of MDR-TB (p value=0.03) Conclusion: Pediatric drug-resistant TB is a rising concern. Awareness programs on national and international levels are needed to educate the general population regarding the importance of preventing TB household contact, especially amongst children.

scholarly journals Uji Diagnostik Genexpert Mtb/Rif Di Rumah Sakit Umum Pusat Haji Adam Malik Medan

2016 ◽  
Vol 1 (2) ◽  
pp. 19
Author(s):  
Elva Susanty ◽  
Zainuddin Amir ◽  
Parluhutan Siagian ◽  
Rina Yunita ◽  
Putri Chairani Eyanoer
Keyword(s):  
Drug Sensitivity ◽  
Surrogate Marker ◽  
Multidrug Resistant ◽  
Sensitivity Testing ◽  
Standard Drug ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis ◽  
Mdr Tb ◽  
Lowenstein Jensen ◽  
Proportion Method

Background: Cases of multidrug resistant tuberculosis (MDR TB) is increasing in number in the world and requires early detection to prevent further transmission. GeneXpert MTB/RIF is a tool that can be used for detection of rifampicin resistance, as a surrogate marker for MDR TB. This study aims to assess the sensitivity and specificity of the GeneXpert MTB/RIF in diagnosis of MDR TB. Methods: diagnostic test study was conducted at a poly MDR TB General Hospital Haji Adam Malik Medan. The subjects were all suspected MDR TB who had results positive GeneXpert MTB/ RIF with sensitive rifampin or resistant  rifampin and had a drug sensitivity test results with the proportion method Lowenstein Jensen medium. Data retrieved from the medical records, between January until December 2013. Results: founded 64 samples that had results of GeneXpert MTB/RIF test positive and had the results of drug sensitivity, 87.5% of rifampin-resistant samples were also resistant to isoniazid. The GeneXpert MTB/RIF examination showed the sensitivity of 92.86% and the specificitu of 59.09%. Conclusion: GeneXpert MTB/ RIF has a high sensitivity for diagnosing MDR TB compared the gold standard drug sensitivity testing proportion method on Lowenstein Jensen medium. This study recommends the GeneXpert MTB/RIF be used for MDR TB screening tool. Keywords: GeneXpert MTB/RIF, multidrug resistant tuberculosis, drug susceptibility test, Lowenstein Jensen medium

scholarly journals Burden of drug-resistant pulmonary tuberculosis in Pakistani children: A cross-sectional study

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 344
Author(s):  
Ghulam Shabbir Laghari ◽  
Zahid Hussain ◽  
Lavina Khemani ◽  
Syed Zohaib Maroof Hussain ◽  
Uzair Yaqoob
Keyword(s):  
Drug Sensitivity ◽  
Sputum Smear ◽  
P Value ◽  
Drug Resistant ◽  
Duration Of Treatment ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Resistant Tuberculosis ◽  
Mdr Tb ◽  
History Of

Introduction: The incidence of drug-resistant tuberculosis (TB) is rapidly increasing worldwide. Children in high TB burden countries are rapidly being reported to be affected by Mycobacterium tuberculosis resistant to isoniazid and rifampicin. The aim of this study is to evaluate the pattern of drug sensitivity among children suffering with TB. Methods: Known cases of pulmonary TB, with sputum smear positive for acid-fast bacilli after two months of compliance to 1st line anti-tuberculous therapy were included after gaining informed consent. Specimens used for drug sensitivity testing were either sputum or bronchoalveolar lavage. Patient age, gender, history of TB contact, and duration of treatment were also recorded. Data was entered and analyzed using SPSS v.22. Results: Fifty children, 32 male (64%) and 18 female (36%) were included in the study. Their mean age was 12.84 ± 2.54 years. History of household TB contact was positive in 29 (58%) children. Among 1st line anti-tuberculous therapy, rifampicin resistance was highest at 33/50 (66%), and resistance to streptomycin and ethambutol were the lowest (6/50; 12%). There were 18 (36%) children with multidrug-resistant tuberculosis (MDR-TB). A positive history of household TB contact (either resistant or non-resistant) was seen to have a statistically significant impact on incidence of MDR-TB (p value=0.03) Conclusion: Pediatric drug-resistant TB is a rising concern. Awareness programs on national and international levels are needed to educate general population regarding the importance of preventing TB household contact, especially amongst children. With the selected method used to identify mainly older children with drug resistance, the yield for drug-resistant TB was found to be high.

scholarly journals Spectral and Antibiotic Susceptibility of Pathogens Isolated from Saudi Patients with Diabetic Foot Infections

2021 ◽  
Vol 12 (1) ◽  
pp. 16-20
Author(s):  
Samiah Hamad S Al-Mijalli
Keyword(s):  
Staphylococcus Aureus ◽  
Diabetic Foot ◽  
Antibiotic Susceptibility ◽  
Empirical Therapy ◽  
Multidrug Resistant ◽  
Foot Ulcers ◽  
Sensitivity Testing ◽  
Cross Sectional ◽  
Diabetic Foot Infections ◽  
Microbiological Methods

Diabetic foot infections (DFIs) are a significant health issue and a common complication among patients with diabetes. To develop antibiotic therapy for these high-risk patients, the current study evaluates the scope of DFIs and identifies the causing microbes. It also measures spectrum and antibiotic susceptibility of the pathogens isolated from adults with DFIs in Saudi Arabia. To achieve the study objectives, a cross-sectional study was implemented and the baseline characteristics for 44 patients with DFIs were defined. Optimal aerobic and anaerobic microbiological techniques were utilized to culture specimens isolated from infected foot ulcers. The standard microbiological methods were employed to identify the bacterial isolates and antibiotic susceptibility testing was conducted following the procedures of the Clinical and Laboratory Standards Institute (CLSI). Results showed that 12 microorganisms were isolated from the participants’ diabetic foot ulcers. Staphylococcus Aureus was ranked first because it appeared in 29 (65.9%) cases. Streptococcus Agalactiae was ranked second and multi-microbial infections were also found. Most of the organisms were susceptible to Vancomycin, Ciprofloxacin, and Cefalexin, but they were resistant to Methicillin, Gentamicin, and Ampicillin antibiotics. Staphylococcus Aureus was most sensitive to Ciprofloxacin, while it was resistant to Methicillin. About 10% of the isolates were multidrug-resistant. The study concludes that while Vancomycin should be used empirically for Gram-positive isolates, Ciprofloxacin can be taken into consideration for most of the Gram-negatives aerobes. Based on including various microorganisms and the advent of multidrug-resistant strains, proper culture and sensitivity testing are necessary prior to the empirical therapy.

scholarly journals Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable Mycobacterium tuberculosis

2021 ◽  
Vol 10 (15) ◽  
pp. 3249
Author(s):  
Annelies W. Mesman ◽  
Seung-Hun Baek ◽  
Chuan-Chin Huang ◽  
Young-Mi Kim ◽  
Sang-Nae Cho ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Media ◽  
Multidrug Resistant ◽  
Culture Conditions ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Solid Culture ◽  
Lowenstein Jensen ◽  
Genetic Mechanisms ◽  
Lipid Resuscitation

An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable Mycobacterium tuberculosis (Mtb) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom Mtb could not be grown in standard solid culture media. When we attempted to re-grow Mtb from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of Mtb from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with Mtb’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance.

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