Sputum Mycobacterium Research Articles

Characteristics of smear-positive pulmonary tuberculosis with normal chest radiographic findings during a tuberculosis outbreak in northeast Malaysia: A cross-sectional study.

Tuberculosis (TB) remains a highly prevalent disease in Malaysia. Early identification using cost-effective methods such as chest radiography can reduce the health burden of a TB epidemic. However, normal chest radiographic findings are common. This study aimed to evaluate the characteristics of smear-positive pulmonary TB with normal chest radiographic findings during an outbreak. A cross-sectional study was conducted by reviewing the medical records of 56 pulmonary TB cases registered at Kodiang Health Clinic from April to October 2022. Smear-negative and extrapulmonary TB cases were excluded. Relevant information was extracted from the medical records and recorded in a case report form for data management and analysis. Approximately 60.7% of the cases had symptoms lasting >2 weeks, and 89.3% showed abnormal findings upon clinical examination. Additionally, 73.2% had sputum acid-fast bacilli counts of ≥1+, and the sputum Mycobacterium tuberculosis culture and sensitivity test findings were positive in 82.1% of the cases. The proportion of smear-positive pulmonary TB with normal chest radiographic findings was 42.9%. The factors associated with smear-positive pulmonary TB with normal chest radiographic findings included being under 18 years old (P=0.021), being a student (P=0.010), being single (P=0.012) and being asymptomatic (P=0.04). Normal chest radiographic findings may lead to a misdiagnosis of smear-positive pulmonary TB, especially during an outbreak. Therefore, active case detection among close contacts with risk factors for normal radiographic findings should consider additional supportive tests.

Adjunctive N-Acetylcysteine and Lung Function in Pulmonary Tuberculosis

BackgroundTuberculosis remains a global health concern, and half of cured patients have permanent lung injury. N-acetylcysteine (NAC) has shown beneficial antimicrobial, antioxidant, and immunomodulatory effects in preclinical tuberculosis models. We examined its effects on tuberculosis treatment outcomes.MethodsThis prospective, randomized, controlled trial nested within the TB SEQUEL cohort study enrolled 140 adults with moderate or far-advanced tuberculosis. Participants were randomly assigned 1:1 to standard therapy with or without 1200 mg of oral NAC twice daily for days 1 to 112. Clinical evaluations, sputum culture, and spirometry were performed at specified intervals through day 168, after which participants returned to the TB SEQUEL cohort. The primary outcome was culture conversion. Secondary outcomes included whole-blood glutathione levels and lung function.ResultsParticipants were predominantly young, male, and human immunodeficiency virus 1–negative and had heavy sputum Mycobacterium tuberculosis (MTB) infection burdens. NAC increased glutathione levels (NAC × day interaction, 8.48; 95% confidence interval [CI], 1.93 to 15.02) but did not increase stable culture conversion (hazard ratio, 0.84; 95% CI, 0.59 to 1.20; P=0.33). NAC treatment was associated with improved recovery of lung function (NAC × month, 0.49 [95% CI, 0.02 to 0.95] and 0.42 [95% CI, −0.06 to 0.91] for forced vital capacity and forced expiratory volume in the first second, respectively, as percentages of predicted values). The effects of NAC on lung function were greatest in participants with severe baseline lung impairment and appeared to persist beyond the period of NAC administration. Rates of serious or grade 3 to 4 nonserious adverse events did not differ between the groups.ConclusionsDespite increasing whole-blood glutathione levels, NAC did not affect eradication of MTB infection in adults with pulmonary tuberculosis that was moderate to far advanced. Secondary outcomes of lung function showed changes that merit further investigation. (Funded by TB SEQUEL grant 01KA1613 of the German Ministry for Education and Research, the Health Africa Project, and the German Center for Infection Research; ClinicalTrials.gov number, NCT03702738.)

Elevated Plasma Matrix Metalloproteinase 8 Associates With Sputum Culture Positivity in Pulmonary Tuberculosis.

Current methods for tuberculosis treatment monitoring are suboptimal. We evaluated plasma matrix metalloproteinase (MMP) and procollagen III N-terminal propeptide concentrations before and during tuberculosis treatment as biomarkers. Plasma MMP-1, MMP-8, and MMP-10 concentrations significantly decreased during treatment. Plasma MMP-8 was increased in sputum Mycobacterium tuberculosis culture–positive relative to culture-negative participants, before (median, 4993 pg/mL [interquartile range, 2542–9188] vs 698 [218–4060] pg/mL, respectively; P = .004) and after (3650 [1214–3888] vs 720 [551–1321] pg/mL; P = .008) 6 months of tuberculosis treatment. Consequently, plasma MMP-8 is a potential biomarker to enhance tuberculosis treatment monitoring and screen for possible culture positivity.

Study on the Results of Sputum Acid Fast Bacteria Smear and Mycobacterium Culture in the Detection of Pulmonary Tuberculosis

Study on the Results of Sputum Acid Fast Bacteria Smear and Mycobacterium Culture in the Detection of Pulmonary Tuberculosis

Dynamics of sputum conversion during effective tuberculosis treatment: A systematic review and meta-analysis.

Two weeks' isolation is widely recommended for people commencing treatment for pulmonary tuberculosis (TB). The evidence that this corresponds to clearance of potentially infectious tuberculous mycobacteria in sputum is not well established. This World Health Organization-commissioned review investigated sputum sterilisation dynamics during TB treatment. For the main analysis, 2 systematic literature searches of OvidSP MEDLINE, Embase, and Global Health, and EBSCO CINAHL Plus were conducted to identify studies with data on TB infectiousness (all studies to search date, 1 December 2017) and all randomised controlled trials (RCTs) for drug-susceptible TB (from 1 January 1990 to search date, 20 February 2018). Included articles reported on patients receiving effective treatment for culture-confirmed drug-susceptible pulmonary TB. The outcome of interest was sputum bacteriological conversion: the proportion of patients having converted by a defined time point or a summary measure of time to conversion, assessed by smear or culture. Any study design with 10 or more particpants was considered. Record sifting and data extraction were performed in duplicate. Random effects meta-analyses were performed. A narrative summary additionally describes the results of a systematic search for data evaluating infectiousness from humans to experimental animals (PubMed, all studies to 27 March 2018). Other evidence on duration of infectiousness-including studies reporting on cough dynamics, human tuberculin skin test conversion, or early bactericidal activity of TB treatments-was outside the scope of this review. The literature search was repeated on 22 November 2020, at the request of the editors, to identify studies published after the previous censor date. Four small studies reporting 3 different outcome measures were identified, which included no data that would alter the findings of the review; they are not included in the meta-analyses. Of 5,290 identified records, 44 were included. Twenty-seven (61%) were RCTs and 17 (39%) were cohort studies. Thirteen studies (30%) reported data from Africa, 12 (27%) from Asia, 6 (14%) from South America, 5 (11%) from North America, and 4 (9%) from Europe. Four studies reported data from multiple continents. Summary estimates suggested smear conversion in 9% of patients at 2 weeks (95% CI 3%-24%, 1 single study [N = 1]), and 82% of patients at 2 months of treatment (95% CI 78%-86%, N = 10). Among baseline smear-positive patients, solid culture conversion occurred by 2 weeks in 5% (95% CI 0%-14%, N = 2), increasing to 88% at 2 months (95% CI 84%-92%, N = 20). At equivalent time points, liquid culture conversion was achieved in 3% (95% CI 1%-16%, N = 1) and 59% (95% CI 47%-70%, N = 8). Significant heterogeneity was observed. Further interrogation of the data to explain this heterogeneity was limited by the lack of disaggregation of results, including by factors such as HIV status, baseline smear status, and the presence or absence of lung cavitation. This systematic review found that most patients remained culture positive at 2 weeks of TB treatment, challenging the view that individuals are not infectious after this interval. Culture positivity is, however, only 1 component of infectiousness, with reduced cough frequency and aerosol generation after TB treatment initiation likely to also be important. Studies that integrate our findings with data on cough dynamics could provide a more complete perspective on potential transmission of Mycobacterium tuberculosis by individuals on treatment. Systematic review registration: PROSPERO 85226.

Surgical treatment of children with a combination of congenital lung malformations and tuberculosis

Congenital malformations of the bronchopulmonary system in combination with tuberculosis are extremely rare in practice, but they usually require surgical treatment. This combination often causes diagnostic difficulties.Characteristics of children and research methods. We have analyzed the experience of surgical treatment of a combination of various congenital lung defects and tuberculosis in 20 children. 9 (45%) children had lung sequestration, 4 (20%) children had lung hypoplasia, 5 (25%) children had congenital bronchiectasis, 1 (5%) child had arteriovenous malformation, and 1 (5%) child had a congenital lung cyst. There were 3 segmental resections, 15 lobeand bilobectomies, and 3 pneumonectomies. All surgeries were performed using minimally invasive approaches (VATS).Results. 11 (55%) patients had diagnostic difficulties of concomitant pathology. Only in 5 (25%) cases they found mycobacteria in sputum before surgery. Postoperative study of material from tuberculous lesions helped to establish multidrug resistance of mycobacterium tuberculosis and to change the chemotherapy regimen in 3 (15%) patients. There were no postoperative complications or deaths. All surgeries were effective. Long-term results were studied in 16 (80%) of 20 patients over a period from 1 to 10 years. No cases of reactivation of tuberculosis or disability.Conclusion. Resection surgery for combination of tuberculosis and congenital lung defects allowed to clarify the diagnosis and to achieve a direct effect in all patients by clarifying the drug resistance of mycobacterium tuberculosis and to optimize postoperative chemotherapy.

Level of Mycobacteria in Pre-Treatment Sputum and Susceptibility of M. tuberculosis Strains Isolated from Patients with non-Multidrug-Resistant Pulmonary Tuberculosis AFB (+) and Factors Influencing MGIT Results after the First 8 Weeks of Anti-Tuberculosi

Vietnam is among 30 high TB ​​burden countries even though the Vietnam National TB Program has made great efforts to detect and treat tuberculosis. Objectives: Assessment of Mycobacterial level in sputum before treatment, and susceptibility to the first line anti-TB drugs of M. tuberculosis strains isolated from TB patients with AFB (+) and non-multidrug-resistant. Moreover, factors influencing MGIT outcome after the first 8 weeks of first-line anti-TB drugs therapy in patients with pulmonary tuberculosis was also analysed. Methodology: An observational, analytical study was performed in 128 patients with non-multidrug-resistant pulmonary tuberculosis AFB (+) for evaluating the level of Mycobacteria in sputum before treatment by smear microscopy method; the susceptibility of M. tuberculosis isolated from sputum of the patient was analysed by Lowenstein - Jensen method. Factors affecting positive MGIT results after 2 months of treatment were determined by multivariate logistics regression. Results: The patients had AFB 3+ were 28% in new cases and 24,5% retreatment patients. The rate of M. tuberculosis strains was susceptible to the first line anti-TB drugs in new cases was higher than retreatment patients. The percentage of any anti-TB drug resistance in retreatment tuberculosis was 59,6%, higher than that of new case TB (23,6%). There was high rate of M. tuberculosis strains resistant to Streptomycin and Isoniazid (12,5% and 16,8% for new cases; 42,3% and 36,5% for retreatment cases, respectively). Large radiographic chest lesions and high AFB levels in pre-treatment sputum were factors associated with a positive MGIT result after the first 8 weeks of treatment. Conclusion: Most of TB patients had high level of Mycobacteria in sputum samples collected before treatment. The percentage of M. tuberculosis strains isolated from sputum of pulmonary non MDR-TB patients had any anti-TB drug resistance were high. High Mycobacteria level in pre-treatment sputum and radiographic chest lesions related to positive MGIT result after the first 8 weeks of treatment.
 Keywords
 Pulmonary tuberculosis, first-line anti-TB drugs, anti-TB drug resistance, susceptibility, M. tuberculosis.
 References
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Effectiveness of intravenous isoniazid and ethambutol administration in patients with tuberculosis meningoencephalitis and HIV-infection

Aims and objectives: of our study was to investigate the effectiveness of intravenously (i/v) isoniazid (H) and ethambutol (E) administration in patients with new sputum positive drug-susceptible pulmonary tuberculosis (TB) with tuberculous meningoencephalitis (TM) and human immunodeficiency virus (HIV) co-infection in the intensive phase of treatment. Methods: Fifty-four patients with TB / TM and HIV co-infection were enrolled in this study. Group 1 included 23 patients treated with E and H i/v, while rifampicin and pyrazinamide were prescribed orally. Group 2 consisted of 31 patients treated with the first-line anti-TB drugs orally. Concentrations of H and E in blood serum were detected using a chromatographic method. Results: A significant improvement in clinical symptoms and X-ray signs in patients treated i/v with H and E was observed compared with group 2. Sputum Mycobacterium tuberculosis positivity was observed during the second month of treatment in 25.0 % of patients from group 1 and 76.1% of patients from the control group (p<0.01). In addition, 9 (39.1%) patients died up to six months when H and E were prescribed i/v compared with 22 (70.9%) in group 2 (p=0.02). Conclusions: In TB / TM with HIV, i/v H and E treatment was more effective than oral H and E treatment at 2 months of intensive treatment in sputum conversion as well as in clinical improvement, accompanied by significantly higher mean serum concentrations. In addition, the mortality rate was lower in i/v H and E treatment than oral.

Features of infusion chemotherapy with first-line drugs in patients with tuberculous meningitis and HIV

Objective. To investigate the effectiveness of first-line intravenous drugs (isoniazid and ethambutol) in patients with tuberculous meningitis and HIV during the intensive phase of chemotherapy.
 Materials and methods. 54 patients with newly diagnosed infiltrative pulmonary tuberculosis, tuberculous meningitis, and HIV were included in this study. The presence of Mycobacterium tuberculosis in sputum was observed in all patients. Group 1 consisted of 23 patients receiving ethambutol and isoniazid intravenously, and rifampicin and pyrazinamide were administered orally. Group 2 consisted of 31 patients treated with first-line oral anti-tuberculosis drugs. Serum isoniazid and ethambutol concentrations were determined by chromatographic method.
 Results. There was a greater improvement in clinical symptoms and radiographic evidence in patients receiving intravenous isoniazid and ethambutol compared with group 2. Positivity of sputum Mycobacterium tuberculosis was observed during the second month of treatment in 25.0 % of patients in group 1 and 76.1 % of patients in the control group (p=0.003). In addition, 9 (39.1 %) patients died before 6 months when isoniazid and ethambutol were administered intravenously compared with 22 (70.9 %) in group 2 (p=0.023).
 Conclusions. In tuberculous meningitis and HIV, intravenous treatment with isoniazid and ethambutol was more effective than oral treatment for 2 months of intensive treatment in the form of faster sputum conversion, clinical improvement, and higher mean concentrations of isoniazid and ethambut blood. In addition, mortality was lower with intravenous isoniazid and ethambutol compared with oral treatment.

S1767 Disseminated Mycobacterium avium Complex Ileitis, Duodenitis, and Pancolitis in an Immunocompromised Patient With HIV/AIDS

INTRODUCTION: Patients with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) are at increased risk of opportunistic infections such as Mycobacterium Avium Complex (MAC); this rarely involves the gastrointestinal (GI) tract. Noncompliance with antiretroviral therapy (ART) increases susceptibility. We herein present a young female with HIV/AIDS and disseminated MAC noncompliant with antiretroviral therapy presenting with watery diarrhea and abdominal pain, ultimately diagnosed with MAC duodenitis, ileitis and pancolitis with unique endoscopic findings. CASE DESCRIPTION/METHODS: A 26-year-old female with history of congenital HIV/AIDS on ART (though poorly compliant), disseminated MAC, genital herpes simplex virus (HSV), and oral thrush presented with generalized cramping abdominal pain and watery diarrhea for three weeks. On admission, she was tachycardic and mildly hypotensive. Physical exam revealed a thin female with mild diffuse abdominal tenderness and genital and perianal ulcerations. Labs showed pancytopenia with white blood cell count 2.45 10 × 3 uL, hemoglobin 8 g/dL and platelet count 56 10*3 uL. Absolute CD4 cell count was 2 uL and HIV RNA PCR over 3 million copies/ml. GI PCR panel and Clostridioides difficile testing were negative. Cytomegalovirus PCR was negative. Sputum Mycobacterium PCR confirmed MAC. Computed tomography abdomen/pelvis showed pancolitis without terminal ileum involvement. Treatment was initiated with intravenous piperacillin/tazobactam, micafungin, and acyclovir while continuing azithromycin, ethambutol and rifabutin for disseminated MAC in the setting of HIV/AIDS. Esophagogastroduodenoscopy showed esophageal, gastric and duodenal ulcerations. Colonoscopy showed severely congested mucosa with scattered areas of nodularity and ulceration (Figure 1) throughout the entire colon and examined terminal ileum. Gastric, duodenal, ileal and colonic biopsies were notable for acid-fast bacilli, consistent with known history of disseminated MAC (Figures 2 and 3). Her prognosis remains guarded given comorbidities and overall poor compliance with therapy. DISCUSSION: Although MAC is a common opportunistic infection seen in immunocompromised hosts, colonic and small bowel dissemination has rarely been reported. The diagnosis may be missed, as the clinical manifestations may be nonspecific. This case highlights the importance of including MAC in the differential of an immunocompromised patient presenting with GI manifestations.Figure 1.: Colon with diffuse areas of severely congested mucosa, scattered nodularity, and ulcerations.Figure 2.: Prominent lamina propria expansion by foamy histiocytes within the mucosa and villi of the ileum with notable absence of distended lacteals, characteristic of Mycobacterium Avium Complex (H&E stain, 100x).Figure 3.: AFB stain highlights acid-fast bacilli within foamy macrophages (AFB stain, 600x).

Effectiveness of Intravenous Isoniazid and Ethambutol Administration in Patients with Tuberculosis Meningoencephalitis and HIV Infection.

BackgroundThe aim of this study was to investigate the effectiveness of intravenous isoniazid (H) and ethambutol (E) administered in patients with new sputum positive drug-susceptible pulmonary tuberculosis (TB) with tuberculous meningoencephalitis (TM) and human immunodeficiency virus (HIV) co-infection in the intensive phase of treatment.MethodsFifty-four patients with TB/TM and HIV co-infection were enrolled for this study. Group 1 comprised of 23 patients treated with E and H intravenously, while rifampicin and pyrazinamide were prescribed orally. Group 2 consisted of 31 patients treated with the first-line anti-TB drugs orally. The concentrations of H and E in blood serum were detected using a chromatographic method.ResultsA significant improvement in the clinical symptoms and X-ray signs in patients treated intravenously with H and E was observed and compared to group 2. The sputum Mycobacterium tuberculosis positivity was observed during the second month of the treatment in 25.0% of patients from group 1 and 76.1% of the patients from the control group (p=0.003). In addition, nine patients (39.1%) died up to 6 months when H and E were prescribed intravenously compared with 22 (70.9%) in group 2 (p=0.023).ConclusionIn TB/TM with HIV, the intravenous H and E treatment was more effective than oral H and E treatment at 2 months of intensive treatment in sputum conversion as well as in clinical improvement, accompanied by significantly higher mean serum concentrations. In addition, the mortality rate was lower in intravenous H and E treatment compared to oral treatment.

Histoplasmos in HIV infection patient. Case study

The article presents a case of the development of histoplasmosis in an HIV-infected patient, describes the possibilities of diagnosis and treatment. The causative agent of the disease is Histoplasma capsulatum (dimorphic fungi). Reduced immunity significantly increases the risk of histoplasm infection. Generalized histoplasmosis occurs in HIV-infected patients. Diagnosis of histoplasmosis is very difficult, especially in areas that are not endemic. To confirm the diagnosis, the pathogen is isolated from sputum, scrapings of the oral mucosa, blood, punctate abscesses of the lymph nodes, liver, spleen, bone marrow, smear microscopy, and culture of the fungus.Patient A., born in 1975, went to the doctor with complaints of fever, cough, weakness. On CT Scan of the chest from 12.07.2017, infiltrative focal changes were not detected, mediastinal lymph nodes 3—7 mm. Ultrasound examination of the abdominal cavity: enlarged lymph nodes are determined to 13—15 mm. The CD4 level is 2.5 % 10 cells/μl blood. 13.07.2017, diagnosed: first diagnosed pulmonary and mesenteric lymphatic tuberculosis, Destr.(–), MBT0. Treatment prescribed according to the regimen: isoniazid, rifampicin, ethambutol, pyrazinamide. During treatment, the temperature returned to normal, the condition improved, and on October 23, 2017, the CD4 level was 10 % 197 cells/μl. From 10.10.2017, she receives ART drugs: dolutegravir, emtricitabine, tenofovir. In January, a significant decrease in the mesenteric lymph nodes was observed on ultrasound. 03/27/2018, the patient suddenly had a telado temperature of 39.0 °C, cough, chills. On a radiograph of the chest on April 13, 2018, focal infiltration changes in S1,2 of the right lung, paratracheal lymph nodes measuring 39 × 28 mm, bifurcation — 15 × 8 mm were revealed. The process in the lungs was regarded as tuberculosis of the intrathoracic lymph nodes and lungs. In the sputum Mycobacterium tuberculosis (smear, GeneXpert and culture) were not determined. Treatment: isoniazid, rifampicin, ethambutol, pyrazinamide, levofloxacin and kanamycin. During therapy, the patient's condition worsened. X-ray 7.06.2018: the left lung is transparent. On the right in the upper, middle and lower lobes of the lung S6,9 polymorphic dissemination is noted, in the lower and middle lobes — areas of consolidation. On the right paratracheal lymph nodes up to 45 mm, bronchopulmonary — up to 15 mm. In order to clarify the diagnosis, a thoracoscopy was performed with a biopsy of lung tissue and lymph nodes. Histologically — histoplasmosis of the lungs and lymph nodes. Received itraconazole. After a month of treatment: the temperature is normal, on the radiograph —- a positive dynamics of the process (resorption of focal infiltration changes). For patient was recommended to continue taking itraconazole 200 mg twice a day for 12 months.Сonclusions. In the absence of the effect of anti-tuberculosis therapy in an HIV-infected patient and the absence of bacteria excretion, it is necessary to take into account possible mycotic damage to the lymphatic system and lungs.

Blood transcriptomic markers of Mycobacterium tuberculosis load in sputum.

Peripheral blood transcriptome signatures that distinguish active pulmonary tuberculosis (TB) from control groups have been reported, but correlations of these signatures with sputum mycobacterial load are incompletely defined. We assessed the performance of published TB transcriptomic signatures in Haiti, and identified transcriptomic biomarkers of TB bacterial load in sputum as measured by Xpert® MTB/RIF molecular testing. People in Port au Prince, Haiti, with untreated pulmonary TB (n = 51) formed the study cohort: 19 people with low and 32 with high sputum Mycobacterium tuberculosis load. Peripheral whole blood transcriptomes were generated using RNA sequencing. Twenty of the differentially expressed transcripts in TB vs. no TB were differentially expressed in people with low vs. high sputum mycobacterial loads. The difference between low and high bacterial load groups was independent of radiographic severity. In a published data set of transcriptomic response to anti-tuberculosis treatment, this 20-gene subset was more treatment-responsive at 6 months than the full active TB signature. We identified genes whose transcript levels in the blood distinguish active TB with high vs. low M. tuberculosis loads in the sputum. These transcripts may reveal mechanisms of mycobacterial control of M. tuberculosis during active infection, as well as identifying potential biomarkers for bacterial response to anti-tuberculosis treatment.

Automatic microscopic detection of mycobacteria in sputum: a proof-of-concept

The laboratory diagnosis of lung mycobacterioses including tuberculosis comprises the microscopic examination of sputum smear after appropriate staining such as Ziehl-Neelsen staining to observe acid-fast bacilli. This standard procedure is operator-dependant and its sensitivity depends on the duration of observation. We developed and evaluated an operator-independent microscopic examination of sputum smears for the automated detection and enumeration of acid-fast bacilli using a ZEISS Axio Scan.Z1 microscope. The sensitivity, specificity, positive predictive value, negative predictive values and accuracy were calculated using standard formulations by comparison with standard microscopic examination. After in-house parameterization of the automatic microscope and counting software, the limit of detection evaluated by seeding negative sputa with Mycobacterium bovis BCG or Mycobacterium tuberculosis H37Rv (100–105 bacilli/mL) was of 102 bacilli/mL of sputum with a 100% positivity rate. Then, the evaluation of 93 sputum specimens including 34 smear-positive and 59 smear-negative specimens yielded a sensitivity of 97.06% [84.67–99.93%], a specificity of 86.44% [73.01–92.78%]. Up to 100 smear slides could be stocked for reading in the microscope magazine and results are exportable into the laboratory information system. Based on these preliminary results, we are implanting this automatic protocol in the routine workflow so that only smears detected positive by automatic microscopy are confirmed by standard microscopic examination.

Rapid method for detecting and differentiating Mycobacterium tuberculosis complex and non-tuberculous mycobacteria in sputum by fluorescence in situ hybridization with DNA probes

ObjectiveIn resource-limited tuberculosis-endemic countries, Mycobacterium tuberculosis in sputum is mainly detected by acid-fast bacillus (AFB) staining and the identification of sputum-derived cultures. PCR techniques are practical only in well-resourced laboratories. This study investigated the application of a rapid, simple, and inexpensive fluorescence in situ hybridization (FISH) assay to identify and differentiate M. tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM) in sputum. MethodsThe Mycobacterium/Nocardia Genus (MN Genus)-MTBC FISH assay performed in this study utilizes two different DNA probes labeled with different fluorescent molecules that hybridize respectively with 16S rRNA of the genus Mycobacterium and 23S rRNA of MTBC. The assay was tested on 202 patient sputum samples in Mangaluru, Karnataka State, India. Sputa were first liquefied and bacteria concentrated before performing the FISH assay and parallel culturing and AFB staining. The identities of cultured bacteria from DNA sequencing were compared with FISH assay findings from corresponding sputa. ResultsOf the 202 sputum samples tested, 67 reacted with both MN Genus-specific and MTBC-specific probes, none reacted only with the MTBC-specific probe, and 22 reacted only with the MN Genus-specific probe. The FISH assay yielded results in 2h and had a limit of detection of 2.2×104CFU/ml in sputum spiked with cultured M. tuberculosis. The diagnostic sensitivity, specificity, and positive and negative predictive values of the FISH assay for MTBC in patient sputa were 89.7%, 95.5%, 88.0%, and 92.6%, respectively. NTM were a significant cause of tuberculosis-like infections in Mangaluru. ConclusionsThe MN Genus-MTBC dual probe fluorescence FISH assay previously applied to cultures can also be utilized in resource-limited tuberculosis-endemic countries for rapidly identifying and differentiating MTBC and NTM in sputum samples.

Transmission of Mycobacterium tuberculosis From Patients Who Are Nucleic Acid Amplification Test Negative.

Among adults with signs and symptoms of pulmonary tuberculosis (TB), recognition of transmissible TB has implications for airborne infection isolation and public health activities. Sputum smear-negative TB patients account for around one-fifth of tuberculosis transmission. The tuberculosis transmission risk of TB patients with negative results on nucleic acid amplification test (NAAT) of respiratory specimens has not been established. We sought to estimate the tuberculosis transmission risk of NAAT-negative TB patients. We retrospectively reviewed Maryland TB program data collected from 2004 to 2009, during which time NAAT using the Mycobacterium Tuberculosis Direct Test (MTD) was performed routinely. Patients with sputum Mycobacterium tuberculosis (M.tb) isolates having matching genotypes were assigned to clusters. Transmission sequence was approximated by collection order of individuals' first culture-positive specimens. Minimum transmission risks of NAAT (MTD)-negative TB patients and of smear-negative TB patients were estimated based on individuals' positions within clusters. Among 809 patients with culture-confirmed TB, M.tb genotypes were available for 782 (96.7%). For NAA-negative TB patients, the minimum transmission risk estimate was 5.1% (95% CI 0-11.4). For smear-negative TB patients, the minimum transmission risk estimate was 11.2% (95% CI 7.2-15.3). Minimum transmission risk of NAAT-negative TB patients was lower than that of smear-negative TB patients. However, transmission risk of NAA-negative TB patients appears to not be negligible.

Detection by death: A rapid way to detect viable slow-growing microorganisms achieved using microchannel Electrical Impedance Spectroscopy (m-EIS)

Based on the insight that only living organisms can be killed (and that killing can proceed much faster than cell-growth), we present an approach for the detection of viable microorganisms that is much faster than currently used culture-based methods. We do so by using microchannel Electrical Impedance Spectroscopy (m-EIS) for real-time detection of cell-death on exposure to a killing-agent. m-EIS relies on the fact that when living-cells with non-zero membrane potentials are exposed to high-frequency AC-field, induced-charges accumulate at the membrane-interface. Cell-death is accompanied by a loss of membrane-potential, and hence charge-storage (capacitance). A proof-of-principle for a clinical-application (detection of living mycobacteria in sputum) is demonstrated. Mycobacterium smegmatis (doubling-time ∼3 hours) and Mycobacterium bovis BCG (doubling-time ∼20 hours) in artificial-sputum are both detected in [Formula: see text]3 hours when exposed to amikacin. Times-to-detection (TTDs) are ∼12 hours and ∼84 hours (3 1/2 days), respectively for culture-based detection using current technologies (BD-MGIT-960[Formula: see text]) for samples containing similar loads of M. smegmatis and M. bovis BCG

Molecular detection of sputum Mycobacterium tuberculosis/rifampicin resistance among presumptive pulmonary tuberculosis cases in Borno state, North-Eastern Nigeria

Background: Drug resistance to anti-tuberculosis (anti-TB) medication and human immunodeficiency virus (HIV) infection undermines global control of TB.Aim: The aim of this study is to determine sputum mycobacteria/rifampicin resistance obtained from Xpert MTB/RIF molecular assay in five health facilities in Borno state.Methods: Records of 5518 presumptive TB cases that presented for care from September 2014 to December 2017 were retrieved from TB registers in this multicentre descriptive study.Results: A total of 5518 pulmonary TB presumptive cases, out of which 5484 were drug-sensitive TB (DSTB) and 34 drug-resistant TB (DRTB). The MTB detection rate was 19.1% and it was higher among DRTB with rate of 41.2% than DSTB of 19.0%. The prevalence of rifampicin resistance was 6.1%, with higher preponderance rate of 78.6% among DRTB cases than 5.1% among DSTB cases. Only 2566 (46.5%) had HIV counselling and testing. The MTB detection rate of 22.1% in HIV-negative patients was significantly higher than 16.5% in HIV patients, P = 0.001, 95% confidence interval (CI) = 2.27–8.93. Conversely, RIF resistance of 7.0% obtained in HIV patients was significantly higher than 4.8% in HIV-negative patients, P = 0.03, 95% CI = 0.06–4.34. Previous TB treatment was significantly associated with RIF resistance, P = 0.000, odd ratio = 1318.1, 95% CI = 302.1–6318.3.Conclusion: GeneXpert is a valuable tool for the detection of both MTB and RIF resistance. It is therefore useful for both management and TB infection prevention and control. Given the observed strong association between previous exposure to anti-TB medication and RIF resistance in this report, we advocate mandatory resistance test for TB patients with previous exposure to TB medication in addition to good adherence to TB medication.

MIXED INFECTION OF MYCOBACTERIUM TUBERCULOSIS AND NONTUBERCULOUS MYCOBACTERIA IN SPUTUM CULTURE AT A SINGLE TERTIARY HOSPITAL

RespirologyVolume 22, Issue S3 p. 135-135 APSR Poster Abstracts MIXED INFECTION OF MYCOBACTERIUM TUBERCULOSIS AND NONTUBERCULOUS MYCOBACTERIA IN SPUTUM CULTURE AT A SINGLE TERTIARY HOSPITAL First published: 23 November 2017 https://doi.org/10.1111/resp.13207_141Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume22, IssueS3Supplement: 22nd Congress of the Asian Pacific Society of Respirology, International Convention Centre, Sydney, Australia, 23–26 November 2017November 2017Pages 135-135 RelatedInformation

Transcriptome analysis of mycobacteria in sputum samples of pulmonary tuberculosis patients.

Pulmonary tuberculosis, the disease caused by Mycobacterium tuberculosis, still retains a top rank among the deadliest communicable diseases. Sputum expectorated during the disease continues to be a primary diagnostic specimen and also serves as a reservoir of bacteria. The expression pattern of mycobacteria in sputum will lead to an insight into bacterial adaptation at the most highly transmissible stage of infection and can also help in identifying newer diagnostic as well as drug targets. Thus, in the present study, a whole genome microarray of Mycobacterium tuberculosis was used to elucidate the transcriptional profile of mycobacteria in the sputum samples of smear positive pulmonary tuberculosis patients. Overall, the mycobacteria in sputum appeared to be in a low energy and low replicative state as compared to in vitro grown log phase M. tb with downregulation of genes involved in ATP synthesis, aerobic respiration and translational machinery. Simultaneously, downregulation was also seen in the genes involved in secretion machinery of mycobacteria along with the downregulation of genes involved in the synthesis of phthiocerol dimycocerosate and phenol glycolipids. In contrast, the majority of the genes which showed an upregulation in sputum mycobacteria were of unknown function. Further identification of these genes may provide new insights into the mycobacterial behavior during this phase of infection and may help in deciphering candidates for development of better diagnostic and drug candidates.