Addressing the drug-resistant tuberculosis challenge through implementing a mixed model of care in Uganda.

Worldwide, Drug-resistant Tuberculosis (DR-TB) remains a big problem; the diagnostic capacity has superseded the clinical management capacity thereby causing ethical challenges. In Sub-Saharan Africa, treatment is either inadequate or lacking and some diagnosed patients are on treatment waiting lists. In Uganda, various health system challenges impeded scale-up of DR-TB care in 2012; only three treatment initiation facilities existed, with only 41 of the estimated 1010 RR-TB/MDR-TB cases enrolled on treatment yet 300 were on the waiting list and there was no DR-TB treatment scale-up plan. To scale up care, the National TB and leprosy Program (NTLP) with partners rolled out a DR-TB mixed model of care. In this paper, we share achievements and outcomes resulting from the implementation of this mixed Model of DR-TB care. Routine NTLP DR-TB program data on treatment initiation site, number of patients enrolled, their demographic characteristics, patient category, disease classification (based on disease site and human immunodeficiency virus (HIV) status), on co-trimoxazole preventive therapy (CPT) and antiretroviral therapy (ART) statuses, culture results, smear results and treatment outcomes (6, 12, and 24 months) from 2012 to 2017 RR-TB/MDR-TB cohorts were collected from all the 15 DR-TB treatment initiation sites and descriptive analysis was done using STATA version 14.2. We presented outcomes as the number of patient backlog cleared, DR-TB initiation sites, RR-TB/DR-TB cumulative patients enrolled, percentage of co-infected patients on the six, twelve interim and 24 months treatment outcomes as per the Uganda NTLP 2016 Programmatic Management of drug-resistant Tuberculosis (PMDT) guidelines (NTLP, 2016). Over the period 2013-2015, the RR-TB/MDR-TB Treatment success rate (TSR) was sustained between 70.1% and 74.1%, a performance that is well above the global TSR average rate of 50%. Additionally, the cure rate increased from 48.8% to 66.8% (P = 0.03). The Uganda DR-TB mixed model of care coupled with early application of continuous improvement approaches, enhanced cohort reviews and use of multi-disciplinary teams allowed for rapid DR-TB program expansion, rapid clearance of patient backlog, attainment of high cumulative enrollment and high treatment success rates. Sustainability of these achievements is needed to further reduce the DR-TB burden in the country. We highly recommend this mixed model of care in settings with similar challenges.

Genetic diversity and drug resistance pattern of Mycobacterium tuberculosis strains isolated from pulmonary tuberculosis patients in the Benishangul Gumuz region and its surroundings, Northwest Ethiopia.

INTRODUCTION: Tuberculosis (TB) remains a major global public health problem and is the leading cause of death from a single bacterium, Mycobacterium tuberculosis (MTB) complex. The emergence and spread of drug-resistant strains aggravate the problem, especially in tuberculosis high burden countries such as Ethiopia. The supposedly high initial cost of laboratory diagnosis coupled with scarce financial resources has limited collection of information about drug resistance patterns and circulating strains in peripheral and emerging regions of Ethiopia. Here, we investigated drug susceptibility and genetic diversity of mycobacterial isolates among pulmonary tuberculosis patients in the Benishangul Gumuz region and its surroundings in northwest Ethiopia. METHODS AND MATERIAL: In a cross-sectional study, 107 consecutive sputum smear-positive pulmonary tuberculosis (PTB) patients diagnosed at two hospitals and seven health centers were enrolled between October 2013 and June 2014. Sputum samples were cultured at Armauer Hansen Research Institute (AHRI) TB laboratory, and drug susceptibility testing (DST) was performed against Isoniazid, Rifampicin, Ethambutol, and Streptomycin using the indirect proportion method. Isolates were characterized using polymerase chain reaction (PCR)based Region of Difference 9 (RD9) testing and spoligotyping. Statistical analysis was performed using Statistical Package for the Social Sciences (SPSS) for Windows version 24.0. RESULTS: Of 107 acid-fast-bacilli (AFB) smear-positive sputum samples collected, 81.3% (87/107) were culture positive. A PCR based RD9 testing revealed that all the 87 isolates were M. tuberculosis. Of these isolates, 16.1% (14/87) resistance to one or more drugs was observed. Isoniazid monoresistance occurred in 6.9% (6/87). Multidrug resistance (MDR) was observed in two isolates (2.3%), one of which was resistant to all the four drugs tested. Spoligotyping revealed that the majority, 61.3% (46/75) of strains could be grouped into ten spoligotype patterns containing two to 11 isolates each while the remaining 38.7% (29/75) were unique. SIT289 (11 isolates) and SIT53 (nine isolates) constituted 43.5% (20/46) among clustered isolates while 29.3% (22/75) were ''New" to the database. The dominant families were T, 37% (28/75), CAS, 16.0% (12/75), and H, 8% (6/75), adding up to 51.3% (46/75) of all isolates identified. CONCLUSION AND RECOMMENDATIONS: The current study indicates a moderate prevalence of MDR TB. However, the observed high monoresistance to Isoniazid, one of the two proxy drugs for MDR-TB, reveals the hidden potential threat fora sudden increase in MDR-TB if resistance to Rifampicin would increase. Clustered spoligotype patterns suggest ongoing active tuberculosis transmission in the area. The results underscore the need for enhanced monitoring of TB drug resistance and epidemiological studies in this and other peripheral regions of the country using robust molecular tools with high discriminatory power such as the Mycobacterial Interspersed Repetitive Units -Variable Number of Tandem Repeats (MIRU-VNTR) typing and whole-genome sequencing (WGS).

Drug-Resistant tuberculosis in Ethiopia: Characteristics of cases in a referral hospital and the implications.

Background: Tuberculosis (TB) programs should design intervention strategies based on the sound knowledge of the existing local epidemiology and sociodemographic characteristics of drug-resistant-TB (DR-TB) cases. The aim of the study was to characterize the pulmonary multidrug-resistant (MDR) and rifampicin-resistant (RR) TB cases enrolled in a referral hospital at Addis Ababa, Ethiopia, called All Africa Leprosy, Tuberculosis, Rehabilitation and Training (ALERT) Hospital. Methods: We conducted a descriptive study based on retrospective review of medical records of 340 pulmonary MDR/RR-TB cases enrolled in ALERT Hospital from November 2011 to December 2016. To characterize the cases, we described the distribution of demographic and clinical characteristics. To compare the distribution of demographic and clinical characteristics between male and female cases, we used Pearson's Chi-squared test. Results: Males accounted for 52.9% of the 340 cases. Nine out of ten cases were in the age group of 15-44 years. Sputum acid-fast bacilli smear-positive and human immunodeficiency virus-coinfected cases constituted 63.7% and 18.1% of cases, respectively. The proportion of new cases increased through the years from nil in 2011 to 21.4% in 2016. Adult males above 24 years constituted more than three quarters (77.2%) of the total male cases, while adult females in this age group constituted 56.9%. The age distribution between male and female cases showed significant differences (P < 0.001). Conclusion: There is age disparity between male and female cases with high impact of MDR/RR-TB on productive adult male population. The transmission potential for DR-TB is also high in the community.

Current Affairs, Future Perspectives of Tuberculosis and Antitubercular Agents.

Tuberculosis (TB) is the major threat for humans from past several decades. Even after advent of several antitubercular drugs, researchers are still struggling for the mycobacterial infections in humans are TB and leprosy. Chronic infections caused by Mycobacterium tuberculosis and Mycobacterium leprae. A particular problem with both of these organisms is that they can survive inside macrophages after phagocytosis, unless these cells are activated by cytokines produced by T-lymphocytes, because of this researchers are not yet succeeded in finding effective treatment on TB. In recent years TB has spread globally and became the major issue for world healthcare organizations. Some compounds like benzothiazinones shown promising activity against mycobacterium, few compounds are in pipeline which may exhibit improved pharmacological effect. Decaprenylphosphoryl-d-ribose 2'-epimerase (DprE1) is the vulnerable target for antitubercular drug discovery. DprE1 is a flavoprotein that along with decaprenylphosphoryl-2-keto-ribose reductase catalyses epimerization of decaprenylphosphoryl-d-ribose to decaprenylphosphoryl-d-arabinose through an intermediate formation of decaprenylphosphoryl-2-keto-ribose. This conversion makes DprE1 a potential drug target. Further research requires to tackle the biggest hurdles in Tuberculosis treatment, i.e. multi drug and extensively drug resistance.

Extended spectrum of antibiotic susceptibility for tuberculosis, Djibouti.

In the Horn of Africa, there is a high prevalence of tuberculosis that is reported to be partly driven by multidrug-resistant (MDR) Mycobacterium tuberculosis strictu sensu strains. We conducted a prospective study to investigate M. tuberculosis complex species causing tuberculosis in Djibouti, and their in vitro susceptibility to standard anti-tuberculous antibiotics in addition to clofazimine, minocycline, chloramphenicol and sulfadiazine. Among the 118 mycobacteria isolates from 118 successive patients with suspected pulmonary tuberculosis, 111 strains of M. tuberculosis, five Mycobacterium canettii, one 'Mycobacterium simulans' and one Mycobacterium kansasii were identified. Drug-susceptibility tests performed on the first 78 isolates yielded nine MDR M. tuberculosis isolates. All isolates were fully susceptible to clofazimine, minocycline and chloramphenicol, and 75 of 78 isolates were susceptible to sulfadiazine. In the Horn of Africa, patients with confirmed pulmonary tuberculosis caused by an in vitro susceptible strain may benefit from anti-leprosy drugs, sulfamides and phenicol antibiotics.

Determination of Lipoprotein Z-Specific IgA in Tuberculosis and Latent Tuberculosis Infection.

Tuberculosis (TB) remains one of the most severe infectious diseases. It is still of paramount importance to establish more accurate, rapid, and efficient diagnostic methods. Since infection with Mycobacterium tuberculosis (M. tb) is largely mediated through the respiratory tract, IgA responses against mycobacterial proteins are worthy of investigation for their potential clinical utility. In this study, the IgA response targeting lipoprotein Z (LppZ) was determined by using a homemade ELISA with plasma of TB patients (N = 125), LTBI individuals (N = 92), healthy controls (HCs) (N = 165), as well as TB patients undergoing anti-TB treatment (N = 9). In parallel the antigen-specific IFN-γ release from PBMCs triggered by LppZ and M. tb-specific ESAT-6 or CFP-10 was detected by using an ELISPOT assay. It was found that the LppZ-specific IgA level was dramatically higher in TB patients than in HCs (p < 0.0001). Compared to that before anti-TB treatment, the LppZ-specific IgA level decreased substantially after 2 months of anti-TB treatment (p = 0.0297) and remained at low levels until the end of the treatment. What is more, pulmonary TB patients exhibited significantly higher LppZ-specific IgA-values than extra-pulmonary TB patients (p = 0.0296). Interestingly, the LppZ-specific IgA-values were negatively correlated to the amounts of IFN-γ released in response to LppZ with statistical significance (r = -0.5806, p = 0.0002). LppZ-specific IgA level was also higher in LTBI individuals than in HCs (p < 0.0001). Additionally there were some PPD+ HC individuals with high LppZ-specific IgA levels but the potential of this assay for identifying leaky LTBI in PPD+ HCs needs to be further investigated through follow-up studies. The sensitivity of detecting TB solely with ESAT-6 or CFP-10-specific IFN-γ release was increased by including the LppZ-specific IgA results, respectively, from 86.11 to 100% and 88.89 to 100%; the sensitivity of screening for LTBI was increased from 80.36 to 83.93% and 57.14 to 69.64%, respectively. The higher LppZ-specific IgA responses in TB and LTBI populations than in controls indicated high immunoreactivity to LppZ upon M. tb infection. Although the assay was not efficient enough for independent application in sero-diagnosis, LppZ-specific IgA might become a complementary biomarker for the improvement of TB and LTBI screening.

Molecular Detection and Differentiation of Mycobacterium Tuberculosis Complex and Non-tuberculous Mycobacterium in the Clinical Specimens by Real Time PCR.

Mycobacteria are subdivided into three groups: the Mycobacterium tuberculosis complex, the non-tuberculous mycobacteria called NTM or MOTT (Mycobacteria Other Than Tuberculosis) and Mycobacterium leprae. Over the past few decades, the incidence of infections caused by NTM has increased world wide. The differentiation of Mycobacterium tuberculosis complex from NTM is of primary importance for infection control and choice of antimicrobial therapy. However, there is so far no report in Bangladesh about the detection of NTM and hence differentiation of MTB and NTM. Neither acid-fast bacilli (AFB) staining nor histopathology can discriminate MTB and NTM. In order to detect and differentiate Mycobacterium tuberculosis complex and NTM we used commercially available LyteStar TB/NTM Real Time PCR kit (Altona Diagnostics, Germany) and analyzed 782 clinical specimens from tuberculosis suspected patients. We have found 49 MTB and 74 NTM positive samples from variety of clinical specimens such as sputum, bronchial lavages, body fluids, tissues, needle aspirates and swabs. Many of the PCR positive specimens were AFB negative on direct microscopic examination thus, indicating strong sensitivity of PCR than AFB staining. This is the first report in the country about detection of NTM and it warrants further elaborate investigation. Moreover, our results showed that multiplex real-time PCR assay is an effective sensitive tool for the rapid identification and differentiation of MTB and NTM directly from clinical specimens.

Susceptibilities of MDR Mycobacterium tuberculosis isolates to unconventional drugs compared with their reported pharmacokinetic/pharmacodynamic parameters.

Background: The second-line drugs recommended to treat drug-resistant TB are toxic, expensive and difficult to procure. Given increasing resistance, the need for additional anti-TB drugs has become more urgent. But new drugs take time to develop and are expensive. Some commercially available drugs have reported anti-mycobacterial activity but are not routinely used because supporting laboratory and clinical evidence is sparse. Methods: We analysed 217 MDR M. tuberculosis isolates including 153 initial isolates from unique patients and 64 isolates from follow-up specimens during the course of treatment. The resazurin microdilution assay was performed to determine MICs of trimethoprim/sulfamethoxazole, mefloquine, thioridazine, clofazimine, amoxicillin/clavulanate, meropenem/clavulanate, nitazoxanide, linezolid and oxyphenbutazone. Isoniazid was used for validation. We calculated the MIC 50 and MIC 90 as the MICs at which growth of 50% and 90% of isolates was inhibited, respectively. Results: The MIC 50 s, in mg/L, for initial isolates were as follows: trimethoprim/sulfamethoxazole, 0.2/4; mefloquine, 8; thioridazine, 4; clofazimine, 0.25; amoxicillin/clavulanate, 16/8; meropenem/clavulanate, 1/2.5; nitazoxanide, 16; linezolid, 0.25; and oxyphenbutazone, 40. The MIC 90 s, in mg/L, for initial isolates were as follows: trimethoprim/sulfamethoxazole, 0.4/8; mefloquine, 8; thioridazine, 8; clofazimine, 0.5; amoxicillin/clavulanate, 32/16; meropenem/clavulanate, 8/2.5; nitazoxanide, 16; linezolid, 0.25; and oxyphenbutazone, 60. By comparison, the MIC 90 of isoniazid was >4 mg/L, as expected. There was no evidence that previous treatment affected susceptibility to any drug. Conclusions: Most drugs demonstrated efficacy against M. tuberculosis . When these MICs are compared with the published pharmacokinetic/pharmacodynamic profiles of the respective drugs in humans, trimethoprim/sulfamethoxazole, meropenem/clavulanate, linezolid, clofazimine and nitazoxanide appear promising and warrant further clinical investigation.

Paleomicrobiology of Leprosy.

The use of paleomicrobiological techniques in leprosy has the potential to assist paleopathologists in many important aspects of their studies on the bones of victims, solving at times diagnostic problems. With Mycobacterium leprae, because of the unique nature of the organism, these techniques can help solve problems of differential diagnosis. In cases of co-infection with Mycobacterium tuberculosis, they can also suggest a cause of death and possibly even trace the migratory patterns of people in antiquity, as well as explain changes in the rates and level of infection within populations in antiquity.

Host Immune Responses Differ between M. africanum- and M. tuberculosis-Infected Patients following Standard Anti-tuberculosis Treatment.

Epidemiological differences exist between Mycobacterium africanum (Maf)- and Mycobacterium tuberculosis (Mtb)-infected patients, but to date, contributing host factors have not been characterised. We analysed clinical outcomes, as well as soluble markers and gene expression profiles in unstimulated, and ESAT6/CFP-10-, whole-Maf- and Mtb-stimulated blood samples of 26 Maf- and 49 Mtb-HIV-negative tuberculosis patients before, and after 2 and 6 months of anti-tuberculosis therapy. Before treatment, both groups had similar clinical parameters, but differed in few cytokines concentration and gene expression profiles. Following treatment the body mass index, skinfold thickness and chest X-ray scores showed greater improvement in the Mtb- compared to Maf-infected patients, after adjusting for age, sex and ethnicity (p = 0.02; 0.04 and 0.007, respectively). In addition, in unstimulated blood, IL-12p70, IL12A and TLR9 were significantly higher in Maf-infected patients, while IL-15, IL-8 and MIP-1α were higher in Mtb-infected patients. Overnight stimulation with ESAT-6/CFP-10 induced significantly higher levels of IFN-γ and TNF-α production, as well as gene expression of CCL4, IL1B and TLR4 in Mtb- compared to Maf-infected patients. Our study confirms differences in clinical features and immune genes expression and concentration of proteins associated with inflammatory processes between Mtb- and Maf-infected patients following anti-tuberculosis treatment These findings have public health implications for treatment regimens, and biomarkers for tuberculosis diagnosis and susceptibility.

M. tuberculosis DNA detection in nasopharyngeal mucosa can precede tuberculosis development in contacts.

BACKGROUND: The nasopharynx is a known gateway for some mycobacterial species such as Mycobacterium bovis and M. leprae. M. tuberculosis can cross lymphoepithelial barriers in vitro, but its ability to colonise the nasopharyngeal mucosa in vivo has not been established. OBJECTIVE: To determine if M. tuberculosis can be transiently detected in nasopharyngeal mucosa of tuberculosis (TB) contacts as a preliminary step in the development of tuberculous infection. DESIGN: Exploratory study conducted among asymptomatic household contacts of pulmonary TB cases. A chest X-ray, QuantiFERON(®) TB-Gold or tuberculin skin test and a bilateral nasopharyngeal swab for Xpert(®) MTB/RIF and mycobacterial culture were performed at baseline and repeated 8-12 weeks later. RESULTS: Eighty-nine contacts were enrolled a median of 9 days after the diagnosis of the index case. At baseline, 29.9% were positive for latent tuberculous infection and one subject (1.1%) had a positive Xpert in the nasopharyngeal swab with a normal chest X-ray, negative QuantiFERON and negative induced sputum. After 12 weeks' follow-up, this subject developed a new cough and upper lobe infiltrates and M. tuberculosis grew in sputum. No other cases of active TB were detected at follow-up. CONCLUSION: The detection of M. tuberculosis DNA in the nasopharyngeal mucosa of contacts is an infrequent event that in this instance preceded the development of pulmonary TB. Its pathogenic role requires further investigation.

Strengthening the Tuberculosis Specimen Referral Network in Uganda: The Role of Public-Private Partnerships.

BACKGROUND: Diagnosis of multidrug-resistant tuberculosis and prompt initiation of effective treatment rely on access to rapid and reliable drug-susceptibility testing. Efficient specimen transport systems and appropriate training on specimen referral contribute to optimal and timely access to tuberculosis diagnostic services. METHODS: With support and technical assistance from a public-private partnership (PPP) between Becton Dickinson and the US President's Emergency Plan for AIDS Relief, the Uganda National TB Reference Laboratory (NTRL) and National TB and Leprosy Program redesigned the tuberculosis specimen transport network and trained healthcare workers with the goal of improving multidrug-resistant tuberculosis detection. RESULTS: Between 2008 and 2011, the PPP mapped 93% of health facilities and trained 724 healthcare and postal staff members covering 72% of districts. Strengthening the tuberculosis specimen referral system increased referrals from presumptive multidrug-resistant tuberculosis cases by >10-fold, with 94% of specimens reaching the NTRL within the established target transport time. CONCLUSIONS: This study demonstrates the potential of PPP collaborations with ministries of health to positively influence patient care by strengthening laboratory systems through increased access to drug-susceptibility testing in Uganda. Ongoing efforts to integrate specimen transport networks will maximize resources and improve patient management.

Primary oral tuberculosis in a patient with lepromatous leprosy: Diagnostic dilemma.

Pulmonary tuberculosis (TB) is the most common form of TB. Primary infection can also affect the pharynx, cervical lymph node, intestine, or oral mucosa. Historically, the observed incidence of concomitant infection with leprosy and TB is high. However, reports of concomitant infection in modern literature remain scarce. Most cases reported in the literature had borderline/lepromatous leprosy and pulmonary tuberculosis. Extrapulmonary tuberculosis is reported in only 3.2% of leprosy cases. To the best of our knowledge, this is the first case report of primary oral tuberculosis of the tongue in a patient with lepromatous leprosy with Type 2 lepra reaction. The patient was referred to Directly Observed Treatment, Short-Course clinic and started on Category I treatment. She received oral prednisolone for lepra reaction, which was subsequently tapered and stopped, however, she continued to receive other antileprotic drugs (thalidomide and clofazimine). The patient's general condition improved and she is on regular follow up.

Treatment Outcomes of Childhood TB in Lagos, Nigeria.

BACKGROUND: : Treatment outcomes of tuberculosis (TB) in children are rarely evaluated by most national TB programmes in sub-Saharan Africa. This study evaluated the treatment outcomes of children treated for TB in Lagos State, Nigeria. METHODS: A retrospective review of programme data of the Lagos state TB and the Leprosy control programme in Nigeria from 1 January 2012 to 31 December 2012. Treatment outcomes were categorized according to the national TB guidelines. RESULTS: A total of 535 cases of childhood TB were notified in 2012, representing 6.3% of the total TB cases notified in Lagos state in 2012. The prevalence of TB/HIV co-infection was 29%. The treatment success rate was 79.2% in TB/HIV-negative children compared with 73.4% in TB/HIV-positive children (p = 0.1268). Children <1 year had the worst treatment outcomes (p < 0.001). CONCLUSION: There is a need to intensify effort at improving notification and treatment outcomes in children.

The discovery, function and development of the variable number tandem repeats in different Mycobacterium species.

The method of genotyping by variable number tandem repeats (VNTRs) facilitates the epidemiological studies of different Mycobacterium species worldwide. Until now, the VNTR method is not fully understood, for example, its discovery, function and classification. The inconsistent nomenclature and terminology of VNTR is especially confusing. In this review, we first describe in detail the VNTRs in Mycobacterium tuberculosis (M. tuberculosis), as this pathogen resulted in more deaths than any other microbial pathogen as well as for which extensive studies of VNTRs were carried out, and then we outline the recent progress of the VNTR-related epidemiological research in several other Mycobacterium species, such as M. abscessus, M. africanum, M. avium, M. bovis, M. canettii, M. caprae, M. intracellulare, M. leprae, M. marinum, M. microti, M. pinnipedii and M. ulcerans from different countries and regions. This article is aimed mainly at the practical notes of VNTR to help the scientists in better understanding and performing this method.

Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.

Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

Diagnóstico diferencial de tuberculosis en base a PCR en lesiones granulomatosas clasificadas histopatológicamente / PCR-based differential diagnosis of tuberculosis in histopathologically classified granulomatous lesions

El diagnóstico diferencial de tuberculosis en tejidos fijados en formalina e incluidos en parafina es necesario porque la morfología de la lesión tuberculosa es variada, hay diversos granulomas clasificados en necrobióticos, tuberculoideos, supurativos, sarcoideo, a cuerpo extraño/crónico inespecífico. Las lesiones granulomatosas ocurren en tuberculosis y también en otras infecciones (hongos, parásitos, brucelosis, lepra) en condiciones tóxicas, alérgicas, autoinmunes, tumores y otras. El diagnóstico histológico no es confirmatorio de tuberculosis y en ausencia de una baciloscopia positiva, se hace necesaria la confirmación molecular para el diagnóstico diferencial. Evaluamos la eficacia de la técnica de PCR para la detección de tuberculosis en tejidos fijados y comparamos esos resultados con la histología del granuloma y la baciloscopia. Analizamos 444 biopsias de diferentes tejidos (ganglios, piel, pleura, pulmón, intestino, tejido óseo, mama y otros) de 5 tipos de granulomas: G1.tuberculoideo con necrosis caseosa; G2.tuberculoideo sin necrosis caseosa; G3. supurativo; G4. sarcoideo l; G5. a cuerpo extraño/inespecífico. Utilizamos dos PCR-IS6110 nested para detección del complejo Mycobacterium tuberculosis y un pan PCR-hsp65 nested para detección de Mycobacterium spp. Los resultados obtenidos muestran que la detección de tuberculosis mediante PCR fue significativamente superior que mediante baciloscopia. G1: PCR 69,6%, baciloscopia 31,3%; G2: PCR 26,8%, baciloscopia 6,1%; G3: PCR 16,7%, baciloscopia 6,7%; G4: PCR 7%, baciloscopia 4%; G5: PCR 6,7%, baciloscopia 0%. Concluimos que el diagnóstico molecular de tuberculosis mediante un PCR robusto adaptado a tejidos fijados es eficaz, rápido, sensible y contribuye a la precisión del diagnóstico diferencial en diferentes tipos de granulomas (AU)

The differential diagnosis of tuberculosis in fixed paraffin embedded-tissues is necessary due to both the diverse morphology of tuberculous lesions and the varying histological types of granulomas (necrobiotic, tuberculoid, suppurative, sarcoidal and foreign body/inespecific). Granulomatous lesions occur in tuberculosis, in other infections (fungal, parasitic, brucelosis, lepra), in toxic, allergic and autoimmune, tumours and in conditions of unknown etiology. Diagnosis of tuberculosis cannot be confirmed by histopathology alone and in absence of a positive acid-fast bacilli (AFB) stain, molecular confirmation of tuberculosis is necessary for a correct differential diagnosis. The aim of our study was to assess PCR efficacy for mycobacterial infection detection in fixed tissues and to correlate those findings with granuloma histology and with AFB staining. We analyzed 444 biopsies from various tissues (lymph nodes, skin, pleura, lung, intestine, bone tissue, breast and others) with 5 granuloma types: G1: with caseous necrosis; G2: without caseous necrosis; G3: suppurative; G4: sarcoidal; G5: chronic/nonspecific. For molecular detection, we used nested PCR-IS6110 for Mycobacterium tuberculosis complex and a nested pan PCR-hsp65 for Mycobacterium sp.. The results obtained demonstrated that PCR was significantly better than AFB stain for tuberculosis detection. G1: PCR 69.6%, AFB staining 31.3%. G2: PCR 26.8%, AFB staining 6.1%; G3: PCR 16.7%, AFB staining 6.7%; G4: PCR 7%, AFB staining 4%. G5: PCR 6.7%, AFB staining 0%. We conclude that molecular diagnosis of tuberculosis using robust PCR-based testing adapted to fixed tissues is a fast, efficient and sensitive method that increases the accuracy of the differential diagnosis of granulomatous lesions (AU)

Compounds for use in the treatment of mycobacterial infections: a patent evaluation (WO2014049107A1).

Tuberculosis is one of the main causes of mortality with 1.5 million deaths a year worldwide. The growing emergence of multi- and extremely resistant strains highlights the urgent need of novel antibiotic strategies. Ethionamide, interfering with the mycobacterial membrane biosynthesis, is used in second-line treatment. This molecule is a prodrug, which requires activation by EthA. The patent described in this evaluation (WO2014049107A1) claimed a new family of molecules and their use as antibiotic treatment against mycobacteria such as Mycobacterium tuberculosis, M. leprae and atypical mycobacteria, either as a single active agent or in combination with antibiotics activable by EthA pathway.

Trend of childhood TB case notification in Lagos, Nigeria, 2011-2014.

BACKGROUND: Childhood tuberculosis (TB) has been neglected by national TB programs in sub-Saharan Africa because of the emphasis on adult smear-positive TB cases. About 80,000 HIV children die from TB, and over 550,000 childhood TB cases occur annually, representing 6% of the global TB burden, making TB an important cause of morbidity and mortality in children. Thus, this study assessed the trend of childhood TB cases notified in Lagos, Nigeria from 2011 to 2014. METHODS: Retrospective data review of childhood TB cases notified to the Lagos State TB and Leprosy Control Programme (LSTBLCP) between January 1, 2011 and December 31, 2014. RESULTS: A total of 2396 children were treated for all forms of TB representing 6.8% of the total 35,305 TB cases notified during the study period. This constituted 1102 (46%) males and 1294 (54%) females. There was a progressive increase in the proportion of children treated for TB from 495 (5.9%) in 2011, 539 (6.4%) in 2012, 682 (7.2%) in 2013 and 680 (7.6%) in 2014. Of the total childhood TB cases notified, 16.3-20% were new sputum pulmonary smear positive; 68.2-74.6% were new sputum pulmonary smear negative; while extra-pulmonary TB accounted for 6.7-10.6%. The case notification rate (CNR) of childhood TB per 100,000 increased from 13.4 in 2011, 14.3 in 2012, 17.7 in 2013 and 17.2 in 2014. CONCLUSION: There was an increase in the case notification rate of TB among children between 2011 and 2014. Efforts should be made to sustain this increasing trend.