Background: Tuberculosis is a major global health problem. Tuberculous lymphadenopathy is a most common form of extrapulmonary tuberculosis (EPTB), constitutes 35% of all cases of EPTB. Due to the paucibacillary nature of specimens, smear microscopy and culture offer low sensitivity. Methods: The aim of the present study was to find the clinicodemographic profiles and comparing the performance of Xpert MTB/RIF, conventional polymerase chain reaction (PCR), mycobacteria growth indicator tube (MGIT) 960, histopathological examination, and clinical follow-up of patients in diagnosing of smear-negative tuberculous lymphadenopathy. Results: A total of 140 clinically suspected cervical tuberculous lymphadenitis cases were enrolled in this study. MGIT-960 culture, conventional PCR, and Xpert MTB/RIF were performed. Most of the patients presented with unilateral (87.14%), single (81.42%), matted (87.85%) lymph nodes, 3 cm–6 cm (52.14%), commonly in the right side (68.02%), and associated lung lesion was found in 12.86% of cases. The detection rates of Mycobacterium tuberculosis complex (MTBC) by Xpert MTB/RIF, conventional PCR, and MGIT were 25.71%, 20.71%, and 17.85%, respectively. Both the tests: Xpert MTB/RIF and PCR, PCR and MGIT, Xpert MTB/RIF and MGIT were positive in 15.71%, 15.71%, and 11.42% of cases, respectively. Most of the patients (74.1%) were cured with 6 months of antitubercular drugs. Conclusion: Clinicians often face the diagnostic dilemmas presented in the study. Individual modalities of the diagnosis are available, but all have drawbacks with varied sensitivity and specificity. Combining the available clinical, radiological, and microbiological modality to reach early diagnosis can go a long way to avoid misdiagnosis and unnecessary delay in treatment, especially in cases, without the pulmonary involvement and fulfilling the aim of National Tuberculosis Control Programme for EPTB cases.

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Cutaneous tuberculosis (TB) has several clinical manifestations. The most common forms include cervical scrofuloderma and plaque type of lupus vulgaris (LV), otherwise the rare cases including inguinal scrofuloderma, ulcerative LV, and acute miliary TB. The highlight of this case series was to report various rare clinical manifestations of cutaneous TB.

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Background: Investigation of DNA methylation in Alu repetitive elements (REs) was shown to be a promising field to explore transcriptional changes in human genome under disease condition. To scrutinize the association between Alu methylation and tuberculosis (TB) disease in children, the difference in Alu DNA methylation level was compared with healthy controls. Methods: Whole-blood genomic DNA from 36 TB-infected children and 32 healthy controls was isolated, and the level of Alu repeat DNA methylation was examined by methylation-specific polymerase chain reaction. Results: The median Alu methylation level in TB patients was 30% (Interquartile range [IQR], 25–30%), whereas in healthy controls, it was 75% (IQR, 50–75%) (P < 0.0001). The median level of DNA methylation of Alu RE in TB cases was significantly lower than healthy controls. Receiver operating characteristic curve analysis showed that the area under the curve for diagnosis was 0.969 (95% confidence interval, 0.936–1) (P < 0.0001), with 100% sensitivity and 84% specificity. Conclusion: Our results point out that detection of Alu DNA methylation in whole-blood DNA may be clinically useful tool for the diagnosis and prognosis of TB disease in children.

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Background: Pleural tuberculosis (TB) diagnosis is sometimes controversial because the microbiologic confirmation ratio is very low in pleural fluid. There are few pediatric pleural TB case series in the literature. Methods: We retrospectively evaluated our TB cases below 18 years of age and extracted pleural TB cases. Results: Seven cases with pleural TB were identified. About 42.9% of the patients had isolated pleural TB whereas 57.1% of the patients had accompanying pulmonary TB. Lymphocytic pleural effusion and increased adenosine deaminase (ADA) (>40 U/L) level are found in 85.7% of the patients. Six patients had uncomplicated effusion (transudate) according to Light's criteria and one had complicated effusion (exudate). Lung decortication was needed in three patients. All patients were given 6 months anti-TB medication and recovered completely. Conclusion: In the lymphocyte-predominant pleural effusion, an increased ADA level highly supported TB disease. The complicated effusion (exudate) in pleural TB is not rule; uncomplicated effusion (transudate) could be seen.

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Background: In this study, we have experimentally investigated the effects of different osmolytes including sucrose, sorbitol, urea, and guanidinium chloride (GdmCl) on the stability and structure of the Mycobacterium tuberculosis pyrazinamidase (PZase). PZase converts pyrazinamide to its active form. Methods: In addition, in order to gain molecular insight into the interactions between osmolytes and PZase, we have conducted 1000-ns molecular dynamics simulations. Results: The results indicated that sucrose and sorbitol increase the stability and compactness of the enzyme, whereas in the presence of urea and GdmCl, PZase loses its stability and compactness. Furthermore, the activity of PZase in the presence of sucrose was more than the other solutions. The energetic analyses imply that the electrostatic and van der Waals interactions are the major factors in the osmolyte–PZase interactions. Sorbitol and sucrose, as protective osmolytes, protect the protein structure by utilizing the van der Waals interaction from denaturation. In addition, urea molecules affect the structure of the protein using the hydrogen bonds and van der Waals interactions. Conclusion: The results show that the most important factor in the denaturing effect of GdmCl is the strong interactions of positively charged guanidinium ions with the aspartate and glutamate residues.

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Background: Sputum culture is limited to centralized facilities. Thus, samples require transportation from peripheral laboratories to these facilities, compromising specimen quality since it is difficult to maintain cold chain. We evaluated OMNIgene SPUTUM Reagent (OMS) for transporting sputum samples for tuberculosis (TB) testing. The study was carried out at Noguchi Memorial Institute for Medical Research using sputa from Korle Bu Teaching Hospital and La General Hospital in Ghana. Methods: In a laboratory-based controlled experiment (CE), sputum contaminants were determined on blood agar before treatment with OMS and N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH). TB testing included smear microscopy, culture, and Xpert MTB/RIF. Afterward, two peripheral laboratories were trained to transport sputum samples with OMS without cold chain. Positivity, negativity, and contamination rates were compared between both methods using Chi-square and Fisher's exact tests. Cohen's Kappa was also used to determine agreements. Results: Among 104 sputum samples analyzed in the CE, 93 (89.4%) had bacterial growth on blood agar before decontamination, while 6 (5.8%) and 5 (4.8%) contaminated after NALC-NaOH and OMS treatment, respectively. Contamination was high with NALC-NaOH (12.8%) than OMS (4.3%) on Lowenstein–Jensen media (P < 0.001), but mycobacterial positivity was comparable: NALC-NaOH of 74.5% and OMS of 78.7%. Smear positivity after NALC-NaOH treatment was 89.4% and OMS was 75.9% (P = 0.491). All except one of the samples tested positive by Xpert MTB/RIF after both treatment. Sixteen samples were evaluated in the field experiment and 81.3% yielded positive culture, and no contamination on LJ was observed. Conclusion: Our findings indicate that OMS works well as a transport and decontaminating reagent of samples for TB testing.

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Background: Effective management of multidrug-resistant tuberculosis (MDR-TB) requires cost-effective and rapid screening of rifampicin (RIF) and isoniazid (INH) resistance. Accordingly, a highly promising high-resolution melting (HRM) analysis was evaluated in the detection of mutation in rpoB, katG gene and inhA promoter region in Mycobacterium tuberculosis isolates. Methods: A total of 143 M. tuberculosis isolates comprising phenotypically confirmed 94 MDR and 49 sensitive isolates were analyzed by HRM following real-time-polymerase chain reaction in comparison to gold standard of targeted DNA sequencing of rpoB, katG gene and inhA promoter region. Results: HRM correctly identified MDR-TB by rapid and accurate detection of predominantly and infrequently occurring specific single nucleotide polymorphism in rpoB, katG gene and inhA promoter region. rpoB HRM showed sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 98% each respectively. Predominantly, S531 L/W (TCG → TTG/TGG) mutation accounted for 68.47% of RIF resistance followed by H526Y/R (13.04%, CAC → TAC/CGC), D516Y/V/G (10.86%, GAC → TAC/GTC/GGC), Q513P (4.34%, CAA → CCA), and one rare mutation at codon position L533A (CTG → CGG). Combined KatG and inhA HRM sensitivity, specificity, PPV, and NPV were 90%, 100%, 100%, and 84.48% respectively and detected frequent mutation at codon position S315T/I/N (70%, AGC → ACC, AGC → ACT, AGC → AAC) and rare mutation at codon position T314P (3.3%, ACC → CCC) and 329 (2.2%, GAC → GCC) of katG gene. In inhA, mutations were recorded at mostly promoter position − 15 (10%, C → T) and infrequently at − 8 (3.3%, T → G, T → C). HRM assay limitation noticed in recognizing silent mutation in rpoB as a mutant, nondetection of infrequent mutation S310A in katG, and the inability of detecting mutation outside the targeted region of investigated genes. Conclusion: HRM may prove to be a vital molecular assay in rapid screening of TB cases for early detection of MDR TB, leading to early evidenced-based initiation of antitubercular treatment that will significantly reduce MDR transmission.

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Tuberculous gummas are an unusual form of tuberculosis (TB) accounting for 1%–2% of all cutaneous TB cases. The aim of this article was to describe the epidemiological, clinical, bacteriological, immunological, and therapeutic features of this form upon a case report and a literature review. Forty-eight case reports were identified through a PubMed and Google Scholar search using the following keywords: “tuberculous gumma” and “metastatic tuberculous abscesse.” Tuberculous gumma can occur at any age. Immunodepression is not a sine qua non condition to the development of the disease. Limbs are the most frequent site for gummas. Tuberculous gummas are associated to another location in only 73% of cases. The most frequent associated locations are lung, nodes, and musculoskeletal apparatus. Mantoux test is negative in 38% of cases. Skin stain is positive in 45% of cases and culture in 85% of cases. Giant epithelioid cell granuloma is present in 82.4% of cases. There is no universally accepted chemotherapy regimen for metastatic tuberculous abscesses. The classic 2RHZE/4RH is highly effective.

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Background: This study aimed to explore the disease severity perceived by people affected with leprosy reporting at leprosy referral center, Purulia, West Bengal, India. Methods: A cross-sectional study was conducted among 358 persons affected by leprosy above the age of 18 and married who were reporting at tertiary leprosy referral hospital, Purulia, West Bengal, India. A semistructured questionnaire was prepared to collect the demographic profile, disease profile, and perceived severity of the disease. Results: Among the participants, 41% of them were female, 60% were aged between 18 and 45 years, 58% were literate, and 40% of the participants had physical disability. The participants had multiple feelings of fear, anxiety and sorrow when first diagnosed as leprosy affected. Majority (69%) of the participants had fear of the disease. A significant association was present among males and females feeling fear of leprosy and the female feared more than male. Conclusion: The present study emphasizes the need for continuous counseling and health education for persons affected with leprosy to avoid or minimize the psychological problems. Periodical screening and counseling will improve their psychosocial well-being and quality of life.

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Background: Zoonotic tuberculosis, commonly referred to as bovine, tuberculosis is mainly caused by Mycobacterium bovis subsp. bovis, a member of the mycobacterium tuberculosis complex (MTBC). Infection in humans resembles that caused by Mycobacterium tuberculosis. Mycobacteria Other Than Tuberculous (MOTTs) are also increasingly being associated with tuberculosis humans but reports of infections in animals are scarce, their zoonotic potential being largely ignored. The study aimed to determine the aetiology and public health importance of mycobacteriosis in slaughter cattle in Kenya. Methods: Routine postmortem meat inspection was performed on a subpopulation of 1000 meat carcasses selected randomly from among 7,564 in a municipal abattoir, between January and July, 2015. Carcasses were examined for tuberculous lesions which were then examined for acid-fast bacilli, (AFB), cultured for isolation of mycobacteria and the isolates characterized by DNA analysis. Results: Of the carcasses examined, 218 (21.8%) had lesions in various parts of the carcasses. Acid-fast bacilli were observed in 63/218 of the lesions and suspected mycobacteria isolated from 35 of them. The isolates were identified as M. fortuitum (12), M. bovis subsp. bovis (3), M. shimoidei (2) M. asciaticum, M. interjectuctum, M. szulgai, M. celatum and M. kansaasii at one (1) each. Thirteen (13) of the isolates could not be speciated. Conclusion: The MOTTs identified in the study have been liked to various types of mycobacteriosis in humans. The study therefore highlights the pathogenic and zoonotic potential of MOTTs, indicating that zoonotic tuberculosis should not be restricted to Mycobacterium bovis subsp.bovis.

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Introduction: Tuberculosis (TB) is a significant cause of morbidity and mortality worldwide. The patient compliance with the long treatment regimens is essential for successful eradication. Pyrazinamide (PZA) shortens these regimens from 9 to 6 months, and therefore, improves treatment completion rates. Although PZA is a first-line medication for the treatment of TB, no simple or reliable assay to determine PZA resistance is yet available. In the presence of PZA, only susceptible Mycobacterium tuberculosis strains release pyrazinoic acid (POA). Therefore, the measurement and quantification of released POA is an indicator of PZA resistance. Methods: Two electrochemical sensors were constructed and tested with alternative working electrodes in conjunction with a portable potentiostat to measure the current produced when a potential difference of 2 V is applied to varying concentrations of POA in controlled solutions. Results: The large (13.2 mm) electrochemical sensor was able to detect POA at a minimum concentration of 40 μM to a statistically significant level (P = 0.0190). Similar graphical trends were obtained when testing the electrochemical sensor in the supernatant of a negative microscopic observation drug susceptibility assay culture, irrespective of the presence of PZA. Conclusion: Inexpensive and reusable electrochemical sensors with a portable potentiostat are a promising tool for the detection of POA, a biomarker of PZA susceptible M. Tuberculosis.

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Background: Antimicrobial resistance has rendered certain species of Mycobacterium difficult to treat clinically, particularly the nontuberculous Mycobacterium and Mycobacterium abscessus. While veterinary medicine and human medicine share many classes of antibiotics, there are several antibiotics which are uniquely licensed to veterinary medicine but not human medicine. It was, therefore, the aim of this study to examine the action of eight veterinary antibiotics to a population of multi- and pan-resistant M. abscessus, isolated from the sputum of patients with cystic fibrosis (CF). Methods: Antibiotic susceptibility studies were performed on human clinical isolates of M. abscessus (n = 16), including 11 smooth isolates, 4 rough isolates, and 1 reference isolate (NCTC 13031), against the following 7 veterinary antibiotics (antibiotic class): apramycin (aminoglycoside), cefovecin (cephalosporin), ceftiofur (cephalosporin), framycetin (aminoglycoside), lincomycin (lincosamide), pirlimycin (lincosamide), and spectinomycin (aminocyclitol). Results: M. abscessus isolates were sensitive (100%) to apramycin and framycetin but resistant (100%) to cefovecin, ceftiofur, lincomycin, pirlimycin, and spectinomycin. Conclusion: This study identified that the veterinary aminoglycosides, apramycin, and framycetin, have in vitro activity against multi-resistant clinical isolates of M. abscessus. Further studies should now compare the activity of these antibiotics against amikacin and the human aminoglycoside, advocated in the treatment of disease in CF patients, to determine if these novel antibiotics have a future role for the development in human medicine with such chronic disease patients.

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Background: The aim of this study was to characterize the drug resistance profile, and the specific lineages of Mycobacterium tuberculosis (MTB) strains isolated from patients with pulmonary TB in the state of Khartoum in Sudan. Methods: Consecutive sputum samples and clinical data were collected from 406 smear-positive TB patients with pulmonary TB in 2007–2009. The samples were cultured, and drug susceptibility testing (DST) was performed using the proportion method (PM) on solid Löwenstein–Jensen medium, and species were identified using biochemical methods at the National Reference Laboratory (NRL) in Khartoum. Extracted deoxyribonucleic acid from a total of 120, 60 suspected multidrug-resistant isolates (MDR), and 60 non-MDR isolates were subsequently sent to the WHO supranational reference laboratory (SRL) in Stockholm at the Public Health Agency of Sweden, for confirmation of the drug resistance profile, examinations by line probe assay (LPA), and molecular epidemiology analysis with Spoligotyping. Results: LPA results correlated 100% for non-MDR and 62% for the suspected MDR strains when compared to the DST results obtained by PM at the NRL. Two strains were initially using the PM identified as MDR-TB but later shown by Hain GenoType Mycobacterium CM/AS to belong Mycobacterium avium complex (Mycobacterium intracelluare). These two strains were excluded from the study material for further analysis. The remaining 58 MDR strains were analyzed using LPA, and 36 strains were confirmed as MDR, 10 as rifampicin monoresistant, and eight as isoniazid-monoresistant. Spoligotyping for all the 118 MTB isolates revealed a total of 115 patterns in which four patterns represented major clusters with a total of 108 (91%) of the strains. The CAS1_Delhi/family was the predominant type and detected in 62 isolates (52%), of which 26 were MDR and 36 were susceptible. It was followed by H3/family with 19 (16%) strains, and 11 Latin American Mediterranean3/family, 16 T2/T1, and two strains each of the Beijing and S lineage. Conclusion: Comparison of DST results obtained using PM and LPA showed 100% agreement for the non-MDR strains but only 62% for the MDR strains. Taking in consideration the time, risk of contamination and the cost of labour to identify MDR TB, the LPA have clear advantages in early detection of MDRTB than the PM. Additionally in this study material Spoligotyping revealed the CAS1 Delhi as the most predominant family. We could not see no major difference in lineages between MDR and non-MDR strains.

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Diaphyseal tuberculosis (TB) is a rare case of the skeletal TB. The following report documents the case of a 52-year-old Moroccan woman with a swelling over the right forearm followed by pulmonary TB under treatment for 3 months. The radiographs of the forearm show a lytic image located at the radius mid-diaphysis. The histopathology confirmed the diagnosis. The patient received surgical drainage with trepanation of the bone. The antibacillary chemotherapy was administered for 6 months. It is, therefore, indispensable to bear in mind the possibility of such atypical presentations of TB when making a rapid and pertinent diagnosis and prescribing the appropriate treatment.

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Chyluria is described as passage of chyle in the urine. It results from the formation of lymphaticorenal fistulae due to various etiologies. The most common implicated cause is filariasis especially in endemic areas. Other uncommon causes include thoracic duct trauma or anatomical abnormalities, pregnancy, malignancy, and tuberculosis. Although tuberculosis has been included as a cause, there are only a few case reports suggesting it as an etiology. Tuberculosis can present in various ways. However, chyluria as initial presentation of tuberculosis has never been reported in the literature. We present, here, an extremely rare case of disseminated tuberculosis involving lung, lymph nodes, liver, intestine, and spine presenting as chyluria. The patient responded dramatically with antituberculous therapy with complete clearing of urine.

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Mycobacterium tuberculosis can cause a myriad of clinical manifestations. We describe a case of a patient with end-stage renal failure, who presented with disseminated tuberculosis over the course of five months, manifesting with a self-resolving mediastinal mass, progressive lymphadenopathy, genitourinary, and musculoskeletal tuberculosis.

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Background: Rapid identification of mycobacteria has been made possible with matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) in recent years. Working with high concentrations of mycobacteria in a PC-3 containment facility makes MALDI-TOF cumbersome and costly. Therefore removing the inactivated isolate's protein extract from the PC-3 facility is needed for efficient identification in a routine PC-2 laboratory. Methods: This work describes a novel chemical and mechanical disruption protein extraction method, which provides reliable MALDI-TOF results from solid and liquid media, while ensuring laboratory safety. Results: When compared to sequencing results, 93.9% of the clinical isolates were identified in LJ media and 89% of the clinical isolates were identified in MGIT media. Conclusion: The MIPE protocol produces a high quality protein extract with improved isolate identification without compromising result turn-around-times or laboratory safety.

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Background: Antimicrobial resistance (AMR) has rendered certain species of Mycobacterium difficult to treat clinically, particularly, the nontuberculous Mycobacterium, Mycobacterium abscessus. This bacterium is emerging in specific disease populations, including amongst cystic fibrosis (CF) patients, where AMR represent a true treatment dilemma. Therefore, any innovation with traditional antimicrobial compounds in spices, which increases the potency of existing conventional antibiotics should be investigated. Methods: M. abscessus isolates (n = 9 multidrug-resistant clinical isolates from CF patients + 1 Reference Strain) were examined for their direct susceptibility to 27 spices, as well as the interactive effect of this spice combination to their susceptibility to amikacin and linezolid antibiotic, with standard disk diffusion assay. Results: Five isolates of M. abscessus (5/10; 50%) failed to grow on the spice enriched medium, which included four clinical isolates and the National Culture Type Collection (NCTC) Reference Strain. Of the remaining five isolates which grew on the spice medium, no cultural phenotypic differences were observed, compared to unsupplemented controls. In the case of both amikacin and linezolid, the zone of inhibition increased with the inclusion of the spices. Initially, all isolates of M. abscessus were fully resistant to linezolid (mean zone of inhibition = 0 mm), and growth was to the edge of the antibiotic disk, whereas when in the presence of spices, large zones of inhibition were observed (mean zone of inhibition = 33.3 mm). With amikacin, the mean zone of inhibition increased from 23.2 mm to 32.0 mm, in the presence of spices. Conclusion: These data suggest that the spices were interacting synergistically with the antibiotics, thus making the antibiotic more potent against the bacteria tested. This study is significant as it demonstrates a positive interaction between spices and the conventional antimycobacterial antibiotics, amikacin, and linezolid. Given the burden of AMR to M. abscessus, particularly in a patient with chronic disease such as CF, any food-related innovation that can help maximize the potency of existing antimycobacterial antibiotics is to be encouraged and developed. The specific mechanism as to how spices increase the potency of such antibiotics with M. abscessus needs to be elucidated, as well as novel food (spice) delivery modalities developed, including novel medicinal foodstuffs or functional foods, that can harness this beneficial effect in vivo to medicine and society.

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