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Diagnosing extrapulmonary tuberculosis (EPTB) is challenging. Immunohistochemistry or immunocytochemistry has been used to diagnose tuberculosis (TB) by detection of MPT64 antigen from various extrapulmonary specimens and has shown good diagnostic performance in our previous studies. The test can distinguish between disease caused by Mycobacterium tuberculosis (Mtb) complex and nontuberculous mycobacteria and can be applied on formalin-fixed paraffin-embedded tissue. As the antibodies previously used were in limited supply, a new batch of polyclonal antibodies was developed for scale-up and evaluated for the first time in this study. Our aim was to assess the diagnostic accuracy of the MPT64 test with reproduced antibodies in the high burden settings of Pakistan and India. Patients were enrolled prospectively. Samples from suspected sites of infection were collected and subjected to histopathologic and/or cytologic evaluation, routine TB diagnostics, GeneXpert MTB/RIF (Xpert), and the MPT64 antigen detection test. Patients were followed until the end of treatment. Based on a composite reference standard (CRS), 556 patients were categorized as TB cases and 175 as non-TB cases. The MPT64 test performed well on biopsies with a sensitivity and specificity of 94% and 75%, respectively, against a CRS. For cytology samples, the sensitivity was low (36%), whereas the specificity was 81%. Overall, the MPT64 test showed higher sensitivity (73%) than Xpert (38%) and Mtb culture (33%). The test performed equally well in adults and children. We found an additive diagnostic value of the MPT64 test in conjunction with histology and molecular tests, increasing the yield for EPTB. In conclusion, immunochemical staining with MPT64 antibodies improves the diagnosis of EPTB in high burden settings and could be a valuable addition to routine diagnostics.
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Mycobacterium tuberculosis , Tuberculose Extrapulmonar , Tuberculose , Adulto , Humanos , Criança , Imuno-Histoquímica , Tuberculose/diagnóstico , Tuberculose/microbiologia , Antígenos de BactériasRESUMO
INTRODUCTION: Lung malignancy is one of the most common neoplasms worldwide. Accurate histology sub-typing and identification of gene mutations in lung tumours are considered important to administer targeted therapy for improved clinical outcome. Our aim is to determine the frequency of EGFR mutation and Programmed death ligand-1 (PD -L1) status of lung malignancies in patients attending a rural hospital in Central India. MATERIALS AND METHODS: Formalin-fixed histology diagnosed lung malignancy (n=99) bronchoscopic/trucut lung biopsies were identified and the tissue blocks and slides were retrieved. Histology typing and staging of the lesions was assessed. PD-L1 expression on biopsy was detected by immunohistochemistry using commercially available primary antibody. PD-L1 expression was assessed and semi-quantified based on the intensity and proportion of tumour cells stained for the marker. EGFR gene mutation at exon19 and 21 was detected by polymerase chain reaction of tissue from paraffin blocks. Final analysis was performed on 87 biopsies for status of EGFR mutation and PD-L1 expression. RESULTS: The average age of lung malignancies patients was 63 years, with a preponderance of males. Advance disease in stage III and stage IV was more common in squamous cell carcinoma as compared to adenocarcinoma (p < 0.01). Mutations at exon 19-21 of the EGFR gene were detected in 7/87 (8%) cases of adenocarcinoma and all of these patients were non-smokers. A total of 52.9% of biopsies showed PD-L1 expression, which was higher in adenocarcinoma patients (p=0.04), smokers (p=0.00), and stage II and III patients (p= 0.00). CONCLUSION: EGFR gene mutations at exon 19 or 21 are seen in lung adenocarcinoma cases. PD-L1 expression was observed in EGFR mutated tissues. Our results should be further validated with large sample size and multicenter clinical data before extrapolation to design immunotherapy strategies.
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Adenocarcinoma , Neoplasias Pulmonares , Masculino , Humanos , Pessoa de Meia-Idade , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Hospitais Rurais , Ligantes , Receptores ErbB/genética , Receptores ErbB/metabolismo , Neoplasias Pulmonares/patologia , Adenocarcinoma/patologia , MutaçãoRESUMO
BACKGROUND: Fine needle aspiration cytology (FNAC) is established as a first line investigation for tuberculous lymphadenitis (TBLA). We aimed to describe the various cytomorphologic features of tuberculosis (TB) on FNAC and their contribution in the diagnostic decision-making in suspected TBLA cases. METHODS: Patients with presumptive TBLA were prospectively enrolled (n = 266) and subjected to routine diagnostic work-up for TB, including FNAC samples, and followed until the end of treatment. Patients were categorized as TB or non-TB cases based on a composite reference standard of which the various cytomorphologic patterns were compared. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy was calculated using cross-tabulation. RESULTS: Fifty-six patients were categorized as bacteriologically confirmed TB, 102 as clinically confirmed TB and 108 as non-TB. The most common cytomorphologic pattern among TB cases (59%) was granulomatous inflammation with necrosis, however, about one-third of tuberculous lymphadenitis patients presented with non-granulomatous inflammation, with 21% showing only necrosis and 13% presenting with a reactive pattern. The overall sensitivity and specificity of FNAC was 85% and 66%, respectively. CONCLUSIONS: We found that about one-third of TBLA patients presented without granulomas on FNA, highlighting the importance of considering TB in a wide spectrum of cytomorphology in a high TB burden setting. Our study supports the use of FNAC as a first-line investigation tool for diagnosing TBLA in a low-resource setting due to its relative simplicity and good sensitivity. However, the low specificity of FNAC, emphasizes the need for a second-tier confirmatory test with improved specificity.
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Background: The global expansion of antibiotic-resistant bacteria is a serious concern and is increasing worldwide in both pathogenic and commensal bacteria. The study determined the correlation between individual child-level antibiotic consumption and antibiotic resistance among the commensal Escherichia coli (E.coli) in a cohort of 125 children in rural Ujjain, India. Methods: During a two-year period between August 2014 and September 2016, stool samples were collected at seven-time points from a cohort of 125 children; aged 1-3. A total of six colonies of E.coli per stool sample were collected for antibiotic susceptibility testing. Antibiotic consumption data was collected during the healthcare-seeking follow-up done during the same period. At each of the seven-time points correlation between antibiotic consumption (Defined Daily Dose-DDD/100 patient-days) and antibiotic resistance (number of resistant isolates) was analyzed independently using the Spearman correlation coefficient. Further, mixed-effects logistic regression models were built to study correlation between child-level consumption of penicillin with the number of E.coli isolates resistant to ampicillin, consumption of cephalosporin with resistance to cefotaxime and ceftazidime, consumption of fluoroquinolones with resistance to nalidixic acid and consumption of cotrimoxazole with resistance to cotrimoxazole. Results: Out of 756 illness episodes reported in 125 children 42% were with antibiotic prescriptions and reported a total antibiotic consumption of 55DDD/100 patient-days. The most common antibiotics used were cefixime (J01DD08;72 DDD/100patient/days) followed by ofloxacin (J01MA01;51DDD/100patient-days), cefpodoxime (J01DD13;38DDD/100patient-days) and amoxicillin (J01CA04;28DDD/100patient-days). The highest percentage of resistance was found to the ampicillin (67%) followed by nalidixic acid (52%) and cefotaxime (44%) and when summarized, more than 90% were resistant to cefotaxime, ceftazidime, and co-trimoxazole in commensal E.coli isolates. The consumption of cephalosporins showed weak positive correlation with the resistance to cefotaxime (Coefficient±SE=0.13 ± 0.09,p<0.001). Conclusion: Our findings showed no correlation between individual-level antibiotic consumption and resistance development in commensal E.coli in a rural community environment.
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OBJECTIVES: To explore the healthcare-seeking pathways, antibiotic prescribing and determine the sociodemographic factors associated with healthcare-seeking behaviour (HSB) of caregivers for common illnesses in under-5 (U-5) children in rural Ujjain, India. STUDY DESIGN: Prospective cohort study. STUDY SETTING AND STUDY SAMPLE: The cohort included 270 U-5 children from selected six villages in rural demographic surveillance site, of the R.D. Gardi Medical College, Ujjain, Madhya Pradesh, India. A community-based cohort was visited two times weekly for over 113 weeks (August 2014 to October 2016) to record the HSB of caregivers using HSB diaries. Sociodemographic information was also solicited. PRIMARY AND SECONDARY OUTCOME MEASURES: Primary outcomes: first point of care, healthcare-seeking pathway and quantify antibiotic prescribing for the common acute illnesses. SECONDARY OUTCOME: HSB risk factors were determined using mixed-effects multinomial logistic regression. RESULTS: A total of 60 228 HSB follow-up time points for 270 children were recorded with a total of 2161 acute illness episodes. The most common illnesses found were respiratory tract infections (RTI) (69%) and gastrointestinal tract infections (8%). No healthcare was sought in 33% of illness episodes, mostly for RTIs. The most common healthcare-seeking pathway was to informal healthcare providers (IHCPs, 49% of illness episodes). The adjusted relative risk for obtaining no treatment, home treatment and treatment by IHCPs was higher for RTIs (aRR=11.54, 1.82 and 1.29, respectively), illiterate mothers (aRR=2.86, 2.38 and 1.93, respectively), and mothers who were homemakers (aRR=2.90, 4.17 and 2.10, respectively). Socioeconomic status was associated with HSB, with the highest aRR for no treatment in the lowest two socioeconomic quintiles (aRR=6.59 and 6.39, respectively). Antibiotics were prescribed in 46% (n=670/1450) illness episodes and the majority (85%, n=572/670) were broad spectrum. CONCLUSION: In our rural cohort for many acute episodes of illnesses, no treatment or home treatment was done, which resulted in overall reduced antibiotic prescribing. The most common healthcare-seeking pathway was to visit IHCPs, which indicates that they are major healthcare providers in rural areas. Most of the antibiotics were prescribed by IHCPs and were commonly prescribed for illnesses where they were not indicated.
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Antibacterianos , População Rural , Antibacterianos/uso terapêutico , Criança , Estudos de Coortes , Atenção à Saúde , Feminino , Humanos , Aceitação pelo Paciente de Cuidados de Saúde , Estudos ProspectivosRESUMO
The world is experiencing crisis of antibiotic resistance not only in pathogenic but also in commensal bacteria. We determine the prevalence of antibiotic resistance in commensal Escherichia coli in young children in rural setting of central India and search for its correlations with demographic and behavioral factors. At seven time points during a period of 2 years we collected stool samples from 125 children; aged 1-3 in a rural area of Madhya Pradesh. We isolated six isolates of E. coli per stool sample and subjected them to antibiotic susceptibility testing. We found resistance to ampicillin, quinolones, cephalosporins, sulfamethoxazole, co-trimoxazole, in at least one isolate from 89% to 100% of children. Extended spectrum beta lactamase producing E. coli were identified in all but one child and multidrug resistance was identified in isolates from all children. Female gender (p = 0.04) and higher wealth (p = 0.03) was significantly correlated with less antibiotic resistance. Thus, the high prevalence of antibiotic resistance in commensal E. coli in rural community from India needs urgent measures to control the growing antibiotic resistance crisis.
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Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli/efeitos dos fármacos , População Rural , Adolescente , Antibacterianos/uso terapêutico , Criança , Pré-Escolar , Escherichia coli/classificação , Escherichia coli/genética , Infecções por Escherichia coli/tratamento farmacológico , Feminino , Genes Bacterianos , Humanos , Índia/epidemiologia , Lactente , Masculino , Testes de Sensibilidade Microbiana , Vigilância da PopulaçãoRESUMO
OBJECTIVE: We aimed to investigate the awareness, health care seeking behavior, and diagnostic delay in extrapulmonary tuberculosis (EPTB) in a resource-constrained setting from Central India. SETTING AND METHOD: Questionnaire based interview of 1220 EPTB patients ≥14 years of age was conducted between July 2004 and August 2012 at Ujjain charitable Hospital, Ujjain, India. RESULTS: Only 15% of patients had ever heard about EPTB and 2-4% knew about its prevention or treatment. Only 12% patients first sought medical advice while 49% patients practiced self-medication, 28% consulted traditional healers and 11% drug store/pharmacy. The median patient delay was 8 weeks (4.6-21.4 weeks). Majority (78%) of patients visited ≥3 health centers. Thirty-eight percent patients first visited any government health facility. Majority (97%) who first visited district and primary public health center were referred to private sector for investigations and 82% patients changed the consultation to private doctor after initial visit to public hospital. The median health system delay was 7 weeks (0.6-16.4 weeks). CONCLUSION: Patients had very poor awareness of EPTB. Patients were referred from public to private sector in search of diagnostic facilities. Improvement of public awareness about EPTB and better public-private partnership may contribute towards reduction in diagnostic delay.
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Antibiotic-resistant bacteria are an escalating grim menace to global public health. Our aim is to phenotype and genotype antibiotic-resistant commensal Escherichia coli (E. coli) from humans, animals, and water from the same community with a 'one-health' approach. The samples were collected from a village belonging to demographic surveillance site of Ruxmaniben Deepchand (R.D.) Gardi Medical College Ujjain, Central India. Commensal coliforms from stool samples from children aged 1-3 years and their environment (animals, drinking water from children's households, common source- and waste-water) were studied for antibiotic susceptibility and plasmid-encoded resistance genes. E. coli isolates from human (n = 127), animal (n = 21), waste- (n = 12), source- (n = 10), and household drinking water (n = 122) carried 70%, 29%, 41%, 30%, and 30% multi-drug resistance, respectively. Extended spectrum beta-lactamase (ESBL) producers were 57% in human and 23% in environmental isolates. Co-resistance was frequent in penicillin, cephalosporin, and quinolone. Antibiotic-resistance genes blaCTX-M-9 and qnrS were most frequent. Group D-type isolates with resistance genes were mainly from humans and wastewater. Colistin resistance, or the mcr-1 gene, was not detected. The frequency of resistance, co-resistance, and resistant genes are high and similar in coliforms from humans and their environment. This emphasizes the need to mitigate antibiotic resistance with a 'one-health' approach.
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Antibacterianos/farmacologia , Resistência Microbiana a Medicamentos/genética , Monitoramento Ambiental , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Fezes/microbiologia , Microbiologia da Água , Animais , Pré-Escolar , Água Potável/microbiologia , Escherichia coli/enzimologia , Feminino , Humanos , Índia , Lactente , Masculino , Testes de Sensibilidade Microbiana , Plasmídeos/genética , População Rural , Águas Residuárias/microbiologia , beta-Lactamases/metabolismoRESUMO
BACKGROUND: Delay in diagnosis is one of the most important factors for the control of tuberculosis (TB) in endemic countries like India. As laboratory diagnosis is the mainstay for identification of active disease, we aim to explore and understand the opinions of medical doctors about the laboratory diagnosis of TB in Ujjain, India. METHODS: Sixteen qualified specialist medical doctors from Ujjain were purposefully selected for the study. Individual interviews with the doctors (13 men and 3 women), were conducted. As one interview could not be completed, data from 15 interviews were analyzed using manifest and latent content analysis. RESULTS: Based on perception of the doctors, the theme; 'challenges and need for the laboratory diagnosis of TB' emerged from the following subthemes: (i) Relationship between basic element of the TB diseases process such as 'Symptoms prior to diagnoses' and 'Clinical characteristics of TB', which were not specific enough to diagnose TB (ii) The prevailing conditions such as lack of explicit diagnostic tools, lead to the doctors using the 'multiple tests' or 'empiric treatment' approach (iii) The doctors proposed that there is a need for access to a rapid, single and simple diagnostic test, and a need for awareness and knowledge of the practitioners regarding specific TB investigations, and early referral to improve the situation at resource-limited settings. CONCLUSION: The medical specialists use a 'multiple test' or 'empiric treatment' approach to diagnose TB. According to the participants, there is a low dependence and uptake of the available laboratory TB investigations by medical practitioners. There is an urgent need to have a specific, simple and reliable test, and a protocol, to improve diagnosis of TB and to prevent development of resistant TB.
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Atitude do Pessoal de Saúde , Necessidades e Demandas de Serviços de Saúde , Padrões de Prática Médica/estatística & dados numéricos , Tuberculose Pulmonar/diagnóstico , Adulto , Técnicas de Laboratório Clínico/normas , Testes Diagnósticos de Rotina/normas , Feminino , Humanos , Índia , Entrevistas como Assunto , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Intra-operative cytology is an important diagnostic tool. It has shown to play an important role especially in the diagnosis of central nervous system tumours. The study was done to assess the feasibility of squash cytology as standalone diagnostic test in setting where frozen section facility is not available. MATERIALS AND METHODS: Total 48 patients with various intracranial lesions were initially enrolled in the study. Patients were investigated by various radio-imaging techniques and routine blood investigations. Forty-one patients were operated at Netaji Subhash Chandra Bose medical college, Jabalpur. Intra-operative squash cytology diagnosis was performed and was correlated with histology diagnosis as gold standard. RESULTS: Out of 41 patients, inflammatory lesions were diagnosed in nine patients while benign lesions [most common neurilemmoma and meningioma] were observed in 21 and malignant lesions [astrocytoma was most common] were diagnosed in 11 patients. Diagnostic accuracy of intra-operative squash cytology irrespective of lesion & site was 95%. We were able to inform about the diagnosis to neurosurgeon in 15 minutes in all cases and within 12 minutes in >85% cases CONCLUSION: Squash smear cytology is reliable and rapid standalone diagnostic method and it can assist for intra-operative decision-making diagnosis of intracranial lesions in resource-limited settings where frozen section facility is not available.
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The aim of the study was to evaluate the diagnostic potential of immunocytochemical staining for the detection of Mycobacterium tuberculosis complex-specific antigen MPT64, in tuberculous lymph node aspirates, cerebrospinal fluid, and effusions from pleura and abdomen. One hundred ninety patients with a diagnosis of tuberculosis (cases) and 80 patients with nontuberculous lesions (controls) were enrolled and differentiated on the basis of clinical features, histology, cytology, clinical biochemistry, Ziehl-Neelsen staining, Lowenstein-Jensen culture, and response to antituberculous therapy. Cervical lymph nodes fine-needle aspirate (n = 150), cerebrospinal fluid (n = 27), pleural fluid (n = 41), and peritoneal fluid (n = 52) were collected and stained with anti-MPT64 and anti-BCG antibodies using immunocytochemistry. Nested-PCR for IS6110 was done for comparison and to calculate the diagnostic indices of the ICC staining. ICC staining with anti-MPT64 was positive in 128/190 (67.4%) tuberculous smears and in 4/80 (5%) control smears thus giving sensitivity of 67.4% and the specificity of 95%, while anti-BCG was positive in 112 (58.9%) tuberculous smears and in 16/80 (20%) control smears with sensitivity of 58.9% and specificity of 80%. When diagnostic validation of ICC was done using PCR as the gold standard, the overall sensitivity, specificity, positive- and negative-predictive values for ICC staining in smears with anti-MPT64 was 96, 96, 95, and 97%, respectively, while the corresponding values for anti-BCG were 87, 88, 86, and 88%. ICC staining using anti-MPT64 represents a robust and simple method for establishing an early etiological diagnosis of M. tuberculosis complex infection in extrapulmonary tuberculosis.
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Antígenos de Bactérias/isolamento & purificação , Imuno-Histoquímica/métodos , Tuberculose/diagnóstico , Adolescente , Adulto , Idoso , Antígenos de Bactérias/líquido cefalorraquidiano , Líquido Ascítico/química , Líquido Ascítico/microbiologia , Biópsia por Agulha Fina , Estudos de Casos e Controles , Contagem de Células , Criança , Pré-Escolar , DNA Bacteriano/genética , Feminino , Humanos , Linfonodos/química , Linfonodos/microbiologia , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Mycobacterium tuberculosis/patogenicidade , Derrame Pleural/microbiologia , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Coloração e Rotulagem/métodos , Fatores de Tempo , Tuberculose/microbiologia , Adulto JovemRESUMO
We have developed and evaluated a polymerase chain reaction (PCR) assay suitable for the detection of Mycobacterium tuberculosis DNA from fine needle aspirate smears of patients with tuberculous lymphadenitis. Air-dried fine needle aspirates of cervical lymph nodes from 98 patients with tuberculous lymphadenitis were studied for cytomorphology, detection of acid fast bacilli by Ziehl-Neelsen staining, culture and nested PCR with IS6110 for mycobacteria on DNA eluted from the dried unstained cytology smear. Twenty aspirate smears with diseases other than tuberculosis were similarly tested as controls. Mycobacterial-DNA was amplified by PCR in 84 (85%) cases and in 1 (5%) control. The mycobacteria could be detected by Ziehl-Neelsen stain and culture in 15 (15.3%) and 24 (24.4%) cases, respectively, whereas both tests were negative in controls. When results were compared with nested PCR on DNA from biopsies from the same case as the gold standard, the sensitivity, specificity, positive and negative predictive values of smear PCR were 85%, 95%, 96%, and 59%, respectively. In conclusion, PCR using dried cytology smear material is feasible and is a simple and sensitive technique for an early and specific diagnosis of M. tuberculosis complex. This is particularly useful when cytology is equivocal and can obviate the use of more invasive procedures.