Severity of TB classified by modified bandim TB scoring associates with the specific sequence of esxA genes in MDR-TB patients

Background: The severity of pulmonary TB and detection of multidrug-resistant (MDR-TB) TB strains as potential causative agents could be crucial for the determination of treatment success. This study aimed to analyze the association between the specific sequences of the full esxA gene from MDR-TB sputum isolates and the severity class of MDR-TB patients.Material and Methods: A total of 98 sputum samples that were suspected to be MDR-TB were collected from Dr. Soetomo, Surabaya, Indonesia, from September to December 2016. A total of 24 isolates from the 98 patients were confirmed to have positive MDR-TB based on the GeneXpert test. MDR-TB isolates were tested using PCR targeting 580 bp encompassing the full esxA gene, and the resulting amplicon was sequenced. The severity class of the pulmonary TB patients was assessed using modified Bandim TB scoring.Results: The patient severity classification resulted in a moderate and severe degree of TB in 38% and a mild degree of TB in 63% of patients. Visualization of the PCR results showed that all MDR-TB samples were positive for the 580 bp band, and the sequence results showed 100% homology with that of the virulent wild-type M. tuberculosis H37Rv (NC_000962.3).Conclusions: In the current study, an association between the characteristics of the full esxA gene and the severity class of MDR-TB patients is yet to be found. However, the homologous sequence of all samples, associated with various degrees of disease severity, possess 100% identity with that of wild-type M. tuberculosis H37Rv. Keywords: Modified Bandim TB scoring; severity of TB disease; full sequence of esxA gene

Resistance Mechanism of Mycobacterium Tuberculosis to Anti-Tuberculosis Drugs nad Multidrug Resistant Tuberculosis

The problem of multidrug resistance is worsening in 2008; WHO's global reposrt on ati-tuberculosis drug resistance ever recorded the highest level of multidrug resistant tuberculosis (MDR-TB) in a general population, with an estimated half a million cases occuring globally. Understanding the emergence and transmission of drug resistant tuberculosis urge to develop and implement the strategic plans on prevention and control drug resistant tuberculosis in line with stop TB partnership's Global plan to Stop TB 2006-2015. Progress in understanding of the drug action and resistance is the important key to develop tuberculosis management program, i.e.: diagnostic strategies and treatment management. The aquisition of resistance on Mycobacterium tuberculosis to anti-tuberculosis drugs derives from randomized and spontaneous chromosomal mutation naturally events, reflects the stepwise accumulation of indivudal mutations in several independent genes; and crucial problems on inadequate traetment create the favorable conditions for the selection of drug resistant Mycobacterium tuberculosis, which develop acquired anti-TB drug resistance and multidrug resistant tuberculosis (MDR-TB); and the transmission to contact persons will multiply primary resistance in a population

Susceptibility of Rifampicin-Isoniazid Resistant Mycobacterium Tuberculosis Isolates Against Levofloxacin

Background: Tuberculosis (TB) is a high burden disease in Indonesia with multidrug-resistant (MDR) TB incidence started to increase. Treatment success of MDR-TB globally was low in number than it was targeted which was especially caused by fluoroquinolone resistance. One of the fluoroquinolone is levofloxacin, an antibiotic that has been widely used irrationally as antimicrobial treatment. Therefore, this study investigated the sensitivity and MBC of MDR Mycobacterium tuberculosis isolates against Levofloxacin. Method: The susceptibility test for MDR-Mycobacterium tuberculosis on levofloxacin by standard method with levofloxacin were on concentrations 0,5 μg/ml, 1 μg/ml, and 2 μg/ml. Sample of 8 strains MDR-Mycobacterium tuberculosis were cultured with each concentrations on Middlebrook 7H9 for 1 week incubation. Next, each of the incubated concentration was subcultured on solid media Middlebrook 7H10 for 3 weeks incubation. Colonized agar plates after 3 weeks incubation were confirmed with acid-fast stain. Results: On MB 7H10 with levofloxacin concentration 2 μg/ml showed bactericidal effect 100% by no MDR Mycobacterium tuberculosis colony grew (0/8) while the MB 7H10 with levofloxacin concentration 1 μg/ml and 0,5 μg/ml showed the bactericidal effect 37,5% and 25% respectively. The colonized agar plate implied that the MDR Mycobacterium tuberculosis with levofloxacin concentration 1 μg/ml (5/8) and 0,5 μg/ml (6/8) grew well. Conclusion: Levofloxacin concentration 2 μg/ml was susceptible on MDR Mycobacterium tuberculosis. The concentration 2 μg/ml of levofloxacin could be considered as MBC

Acid-Fast Bacilli Conversion of Beijing and Non-Beijing Strain of Pulmonary Tuberculosis In South Sulawesi

Beijing strains are a major part of the Mycobacterium tuberculosis Asian phylogenetic lineage. Beijing strains represent about 50% of all TB strains in East Asia and at least 13% of strains worldwide. Beijing strain of Mycobacterium tuberculosis is presumed as the factor of the increase in bacteria virulence and drug resistance, and the contributor in treatment failure. The aim of this study was to analyze the association between acid-fast bacilli conversion with strain genotipe Beijing and non-Beijing of pulmonary tuberculosis in South Sulawesi. The design of research was observational analytic with prospective approach. The sampling technique used consecutive sampling. Data were taken from active pulmonary tuberculosis patients’ medical record in Balai Besar Kesehatan Paru Masyarakat Makassar (Pulmonary Health Center of Makassar) and Community Health Center in Gowa Regency, South Sulawesi from March to June 2018. Collected sputum samples were screened for AFB and identified as Beijing strain and non Beijing strains using Multiplex PCR in Tropical Disease Institute of Universitas Airlangga. The results is showed that the characteristics of the respondents consisted of 12 respondents (33.3%) aged 56-65 years, 25 respondents (69.4%) men and 28 respondents (77.8%) had low category gradation of AFB smear. Univariate analysis showed 6 respondents (16.7%) with Beijing strains, 30 respondents (83.3%) with non-Beijing strains, 32 respondents (88.9%) conversion sputum AFB and 4 respondents (11.1%) non conversion sputum AFB. Bivariate analysis with Chi-Square statistical test shows that p value 0.022 < 0,05, that means there was association of Beijing strains with BTA conversion. Microscopic examination of BTA can be used to monitor and evaluate the treatment of new pulmonary TB patients undergoing treatment and the Beijing Mycobacterium tuberculosis strain has a significant correlation with the treatment failure of anti-tuberculosis drugs in South Sulawesi

Nucleic Acid Amplification of The rpoB Region of Mycobacterium tuberculosis in Pulmonary Tuberculosis Diagnosis

Tuberculosis (TB) is one of the major public health concerns worldwide. The detection of the pathogen Mycobacterium tuberculosis complex (MTBC) as early as possible has a great impact on the effective control of the spread of the disease. It is difficult to diagnose Mycobacterium tuberculosis infection due to a lack of rapid, sensitive and specific test. Newer methods, which are easy and reliable, are required to diagnose TB. This research aim is to evaluate the accuracy polymerase chain reaction (PCR) technique, using primers the rpoB gene region compare to culture method in Lowenstein-Jensen medium as a gold standard for the detection of Mycobacterium tuberculosis in the sputum samples. Sputum samples from TB suspected patients are examined by culture and PCR, using rpoB target gene. Specimens are digested and decontaminated by the modified Petroff method (WHO). Approximately from 1.0 ml of resuspended sediment, each 100 ul is used to inoculate Lowenstein-Jensen slants in duplo and 100 ul resuspended sediment is processes for PCR. Mycobacterium tuberculosis is identified using a specific pair of primers designed to amplify 541 bp sequences of rpoB gene. Conclusion: PCR have the high accuracy, sensitivity 100% and specificity 100% for pulmonary TB diagnosis. The performance of a rpoB Mycobacterium tuberculosis PCR assay have value in the rapid diagnosis of pulmonary tuberculosis

Antibacterial activity of probiotic mixed culture against MRSA and ESBL

Faced with the methicilin resistant Staphilococcus aureus (MRSA) and the extended-spectrum beta-lactamase (ESBL) pandemic, this study reports the antibacterial activity of seven probiotics against two MRSA (Px1 and Px2) and six ESBL (Px3, Px4, Px5, Px6, Px7, and Px8) isolated from clinical patients. The mixed culture of probiotics was consisted of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Bifidobacterium bifidum, Bifidobacterium animalis, Lactobacillus plantarum, Streptococcus thermophilus. The probiotics mixture was performed by mixing equally each probiotic starter in MRS broth medium and then incubated at 35oC, 150 rpm for 48 hours. The anti bacterial activity of free cell supernatant (FCS) against each of the bacterial test was assayed by agar diffusion method by using hole as reservoir. The result showed that all the isolates exhibited very good inhibitory activity against target bacteria as indicated by the diameter of inhibition zone: ESBL (21-22 mm) and MRSA (19-20 mm). The potency ratio of the probiotics mixture was also evaluated against kanamycin, streptomycin, and ultra broad spectrum meropenem as standard

Prospects of red passion fruit seeds (Passiflora edulis Sims.) as a source of halal probiotics

Indonesia is the most biodiversity country in the world, with a majority Muslim population, so halal issues are a priority. Natural resources are abundant, especially the tropical fruit that Allah SWT bestowed with a wide variety. Red Passion fruit (Passiflora edulis Sims.) is one of the tropical fruits widely consumed because of its delicious taste, rich in vitamins, minerals, and other active ingredients. The research reported that probiotics of Bacillus spp. Isolated from the red passion fruit seeds was proven to inhibit pathogenic microbes. Extended Strain Beta-Lactamase and Methicillin Resistant Staphylococcus aureus. The isolates also showed the ability to increase gamma interferon levels in the peripheral blood mononuclear cells in adult TB patients. Halal critical points for products containing probiotics can derive from raw material sources, especially those from animals. The Red passion fruit is classified as a safe, halal category. The process of isolating probiotics from red passion fruit seeds using fermentation technology in De man Rogose Sharp media, using simple materials, tools, and steps that are guaranteed to be halal. The compatibility of two probiotic isolates is the potential to be developed into a probiotic consortium for health supplement preparations and therapeutic complements

Risk factors of latent tuberculosis infection in healthcare workers at hospitals in Jember City Indonesia

Background: Healthcare workers in Tuberculosis (TB) and non-TB units in hospitals have a high risk of experiencing Latent Tuberculosis Infection (LTBI), because of exposure to droplets containing Mycobacterium tuberculosis. This study aims to prove LTBI incidence and risk factors to healthcare workers at the hospital in Jember City.Material and Methods: a cross-sectional study, from January to March 2020 in two hospitals in Jember City. Healthcare workers in the TB care and non-TB care unit were examined using Tuberculin skin test (TST) with a cut off ≥ 10 mm for positive LTBI. Chest x-ray and clinical examination to rule out active TB and a standardized questionnaire were also used.Results: 128 healthcare workers completed the questionnaires, clinical, tuberculin skin test (TST), and chest x-ray data. LTBI incidence of positive results 61.7% (n = 79). Contacts TB in the workplace (p value = 0.219; OR = 1.643; CI = 0.742-3.641) and a unit of work (p value = 0.102; OR = 0.760; CI = 0.559-1.031) has no relationship with LTBI. The profession (p value = 0.020; OR = 1.112; CI = 0.896-1.403), the duration of the work (p value = 0.039; OR = 2.984; CI = 1.067-8.342), and BCG immunization (p value =0.000; OR = 0.151; CI = 0.052-0.438) have important relationships with LTBI.Conclusion: TB infection with a high incidence, a risk of transmission to healthcare workers, and a relationship between occupational risk factors and LTBI among healthcare workers in Jember City, Indonesia have been established in this study

RISK FACTORS OF LATENT TUBERCULOSIS INFECTION IN HEALTHCARE WORKERS AT HOSPITALS IN JEMBER CITY INDONESIA

Background:&nbsp;Healthcare workers in Tuberculosis (TB) and non-TB units in hospitals have a high risk of experiencing Latent Tuberculosis Infection (LTBI), because of exposure to droplets containing&nbsp;Mycobacterium tuberculosis.&nbsp;This study aims to prove LTBI incidence and risk factors to healthcare workers at the hospital in Jember City. Material and Methods:&nbsp;a&nbsp;cross-sectional study,&nbsp;from January to March 2020 in two hospitals in Jember City. Healthcare workers in the TB care and non-TB care unit were examined using Tuberculin skin test (TST)&nbsp;with a cut off ≥ 10 mm for positive LTBI. Chest x-ray and clinical examination to rule out active TB and a standardized questionnaire were also used. Results:&nbsp;128 healthcare workers completed the questionnaires, clinical, tuberculin skin test (TST), and chest x-ray&nbsp;data. LTBI incidence of positive results 61.7% (n = 79). Contacts TB in the workplace (p value = 0.219; OR = 1.643; CI = 0.742-3.641) and a unit of work (p value = 0.102; OR = 0.760; CI = 0.559-1.031) has no relationship with LTBI. The profession (p value = 0.020; OR = 1.112; CI = 0.896-1.403), the duration of the work (p value = 0.039; OR = 2.984; CI = 1.067-8.342), and BCG immunization (p value =0.000; OR = 0.151; CI = 0.052-0.438) have important relationships with LTBI. Conclusion:&nbsp;TB infection with a high incidence, a risk of transmission to healthcare workers, and a relationship between occupational risk factors and LTBI among healthcare workers in Jember City, Indonesia have been established in this study. &nbsp; &nbsp; &nbsp; &nbsp; &nbsp

Rapid Culture Method Using Biphasic Media On Bacteriologic Diagnosis To Detect Mycobacterium tuberculosis For Determining Pulmonary Tuberculosis

Diagnosis to determine pulmonary TB based on detection of Mycobacterium tuberculosis rapidly and accurately is important todetermine the adequate therapy. Standard culture method using Lowenstein Jensen (L-J) media to detect and diagnoseMycobacterium tuberculosis require long time. Culture on solid clear medium may increase the sensitivity in examining the growthof mycobacterial colonies. Supplemented nutrition of media with sera in agar media and sera plus blood in broth phase media willincrease growth rate of colonies. Selectivity of culture media by added malachite green and penicillin. The objective was todetermine the validity of culture method using a biphasic blood-sera agar medium comparing with the standard culture method L-Jfor the detection of Mycobacterium tuberculosis from sputum specimens of 100 suspect pulmonary TB. The nutrition composition ofbiphasic media blood-sera agar which optimum colony growth of mycobacteria are 3.3gram nutrient agar (Oxoid) in 90ml aquadest,25μg malachite green, 3ml glycerol, 10ml sera, and 100U/ml penicillin, and broth phase contain similar nutrition by added blooddiluted; these biphasic culture method has a high laboratory sensitivity with limit detection of 102cfu/ml and high specificity forMycobacterium tuberculosis. Growth rate of mycobacterial colonies in biphasic media 10-14 days; biphasic method has high clinicalvalidity, 100% sensitivity and spesificity 100% to detect Mycobacterium tuberculosis in sputum of suspect pulmonary TB. Inconclusion, rapid culture method using biphasic media blood-sera agar for bacteriologic diagnosis to detect Mycobacteriumtuberculosis in sputum of suspect pulmonary TB patients have high sensitivity and specificity