Expert panel consensus recommendations on the utilization of nebulized budesonide for managing asthma and COPD in both stable and exacerbation stages in Thailand.

OBJECTIVE: This study investigated the utilization of nebulized budesonide for acute asthma and COPD exacerbations as well as for maintenance therapy in adults. DATA SOURCES: We conducted a search on PubMed for nebulized budesonide treatment. SELECTED STUDIES: Selecting all English-language papers that utilize Mesh phrases "asthma," "COPD," "budesonide," "nebulized," "adult," "exacerbation," and "maintenance" without temporal restrictions, and narrowing down to clinical research such as RCTs, observational studies, and real-world studies. RESULTS: Analysis of 25 studies was conducted to assess the effectiveness of nebulized budesonide in asthma (n = 10) and COPD (n = 15). The panel in Thailand recommended incorporating nebulized budesonide as an additional or alternative treatment option to the standard of care and systemic corticosteroids (SCS) based on the findings. CONCLUSION: Nebulized budesonide is effective and well-tolerated in treating asthma and COPD, with less systemic adverse effects compared to systemic corticosteroids. High-dose nebulized budesonide can enhance clinical outcomes for severe and mild exacerbations with slow systemic corticosteroid response. Nebulized budesonide can substitute systemic corticosteroids in some situations.

Clusters of Drug-Resistant Mycobacterium tuberculosis Detected by Whole-Genome Sequence Analysis of Nationwide Sample, Thailand, 2014-2017.

Multidrug-resistant tuberculosis (MDR TB), pre-extensively drug-resistant tuberculosis (pre-XDR TB), and extensively drug-resistant tuberculosis (XDR TB) complicate disease control. We analyzed whole-genome sequence data for 579 phenotypically drug-resistant M. tuberculosis isolates (28% of available MDR/pre-XDR and all culturable XDR TB isolates collected in Thailand during 2014-2017). Most isolates were from lineage 2 (n = 482; 83.2%). Cluster analysis revealed that 281/579 isolates (48.5%) formed 89 clusters, including 205 MDR TB, 46 pre-XDR TB, 19 XDR TB, and 11 poly-drug-resistant TB isolates based on genotypic drug resistance. Members of most clusters had the same subset of drug resistance-associated mutations, supporting potential primary resistance in MDR TB (n = 176/205; 85.9%), pre-XDR TB (n = 29/46; 63.0%), and XDR TB (n = 14/19; 73.7%). Thirteen major clades were significantly associated with geography (p<0.001). Clusters of clonal origin contribute greatly to the high prevalence of drug-resistant TB in Thailand.

Microorganisms and clinical outcomes of early- and late-onset ventilator-associated pneumonia at Srinagarind Hospital, a tertiary center in Northeastern Thailand.

BACKGROUND: Ventilator-associated pneumonia (VAP) is a common nocosomial infection in intensive care unit (ICU). Local microbiological surveillance of pathogens and resistance patterns for early-onset VAP (EOVAP) and late-onset VAP (LOVAP) will help to choose appropriate empiric antibiotics. OBJECTIVE: To compare the multi-drug resistant (MDR) pathogens, treatment outcomes, and factors associated with hospital mortality of VAP. METHOD: A cross-sectional study between 1 January 2015 and 31 December 2017 at Srinagarind hospital, Khon Kaen University was conducted. The demographic data, causative pathogens, hospital length of stay (LOS), ICU LOS, mechanical ventilator (MV) days, and hospital mortality were retrospectively reviewed. RESULTS: One hundred and ninety patients were enrolled; 42 patients (22%) were EOVAP and 148 patients (78%) were LOVAP. Acinetobacter baumannii was the most common pathogen in both groups (50% EOVAP vs 52.7% LOVAP). MDR pathogens were significant greater in LOVAP (81.8%) than EOVAP (61.9%) (p = 0.007). The EOVAP had a significantly better ICU LOS [median (interquartile range, IQR) 20.0 (11.0, 30.0) vs. 26.5 (17.0, 43.0) days], hospital LOS [median (IQR) 26.5 (15.0, 44.0) vs. 35.5 (24.0, 56.0) days] shorter MV days [median (IQR) 14.0 (10.0, 29.0) vs. 23.0 (14.0, 35.5) days] and lower hospital mortality (16.7% vs 35.1%) than LOVAP (p < 0.05). The factor associated with hospital mortality was having simplified acute physiology (SAP) II score ≥ 40 with an adjusted odds ratio (aOR) of 2.22 [95% confidence interval (CI), 1.08-4.54, p = 0.02]. CONCLUSION: LOVAP had significantly higher MDR pathogens, MV days, ICU LOS, hospital LOS and hospital mortality than EOVAP. A broad-spectrum antibiotic to cover MDR pathogens should be considered in LOVAP. The factor associated with hospital mortality of VAP was a SAPII score ≥ 40.

The immediate effects of breathing with oscillated inspiratory and expiratory airflows on secretion clearance in intubated patients with cervical spinal cord injury.

STUDY DESIGN: A prospective, randomized crossover trial. OBJECTIVES: To evaluate the efficacy of the combination of incentive spirometry with oscillation (OIS) and positive expiratory pressure with oscillation (OPEP) to promote secretion clearance in intubated patients with cervical spinal cord injury. SETTING: Spinal cord unit, tertiary care hospital, North East Thailand. METHODS: Thirteen intubated patients (C4-7, AIS score C) with secretion retention performed three interventions randomly allocated on consecutive days, a Sham deep breathing, OPEP and OPEP + OIS breathing exercise. Secretions were collected by sterile suction for 3 h before, and 3 h after, each intervention and wet weight recorded. Cardiopulmonary parameters were measured before and after each intervention. RESULTS: The median (IQR) secretion wet weight pre-intervention was 2.61 g (2.21, 3.85) and in the 3 h after Sham there was an increase of 1.97 g (0.6, 3.6). The increase after OPEP was 2.67 g (1.7, 3.9) and after OPEP + OIS, 4.28 g (2.4, 6.7); all the increases being significant (p ≤ 0.007). The clearance after OPEP and OPEP + OIS were both greater than Sham while OPEP + OIS was greater than OPEP (p ≤ 0.019). There were no significant changes in cardiopulmonary measures following any intervention or when compared between interventions. CONCLUSIONS: Deep breathing with an oscillated and humidified air flow in a combination of OIS + OPEP more than doubled secretion clearance and was more effective than OPEP or Sham deep breathing. There were no adverse effects of the procedures which were well tolerated by the patients and may be used to complement existing methods for secretion clearance.

COMPARISON OF GeneXpert MTB/RIF ASSAY WITH CONVENTIONAL AFB SMEAR FOR DIAGNOSIS OF PULMONARY TUBERCULOSIS IN NORTHEASTERN THAILAND.

Among infectious agents, Mycobacterium tuberculosis remains one of the most significant causes of death worldwide. Rapid and accurate diagnosis of pulmonary tuberculosis (TB) remains a great challenge. GeneXpert MTB/RIF assay is a novel integrated diagnostic system for rapid diagnosis of TB and particularly of rifampicin-resistant strains. A study was conducted between January 2010 and December 2014 to compare the performance of the sputum GeneXpert MTB/RIF assay with the conventional sputum AFB smear for diagnosis of active pulmonary TB in Thailand, a country with a high burden of this disease. Of the 125 patients who had cough and/or prolonged fever together with abnormal chest radiograph, 63 were diagnosed as having pulmonary TB by mycobacterium culture assay, while the remaining subjects were considered of having TB-like conditions, viz non-tuberculous mycobacterium infection (NTM), bacterial pneumonia or bronchogenic carcinoma. Two-thirds of the patients had underlying diseases, eg, diabetes mellitus (19 patients), autoimmune diseases (14), and HIV (6). Among patients with positive diagnosis of M. tuberculosis infection, 30 were AFB smear positive and 53 by sputum GeneXpert MTB/RIF method; among patients negative for M. tuberculosis infection, 4 were AFB smear positive and 5 by GeneXpert MTB/ RIF assay. Sensitivity and specificity of the sputum AFB smear and GeneXpertMTB/ RIF assay test were 48% (95% CI: 35-61) and 84% (95% CI: 73-92), and 94% (95% CI: 84-98) and 92% (95% CI: 82-97), respectively. Diagnostic performance of the GeneXpert MTB/RIF assay among AFB smear positive patients was higher than among AFB smear negative patients (adjusted OR 6.7; 95% CI: 2.3-19.9). Earlier diagnosis of pulmonary TB using GeneXpert MTB/RIF assay will lead to earlier appropriate treatment and provide opportunities to interrupt TB transmission.

DIAGNOSTIC TEST OF SPUTUM GENEXPERT MTB/RIF FOR SMEAR NEGATIVE PULMONARY TUBERCULOSIS.

The objective of this study was to evaluate the performance of the Gene-Xpert MTB/RIF sputum test for diagnosing pulmonary tuberculosis (TB) among patients sputum acid-fast bacillus (AFB) smear negative results in Thailand, a country with a high prevalence of pulmonary tuberculosis. We studied 151 patients who presented to Srinagarind Hospital, Khon Kaen, Thailand with a 2 week or more history of fever and/or cough and an abnormal chest radiograph between 2010 and 2014; these patients had at least 2 negative sputum AFB smear results. Of these, 76 were diagnosed as having either confirmed or probable pulmonary TB: the 32 confirmed cases were those with a positive sputum culture for Mycobacterium tuberculosis (MTB) and the 44 probable case were those with clinical and radiographic findings consistent with TB and who had a response to anti-TB therapy. Seventy-five cases were diagnosed as not having pulmonary TB. Of the 32 patients with a positive sputum culture for MTB, 26 had a positive GeneXpert MTB/RIF sputum test. Compared to sputum culture for MTB the GeneXpert MTB/ RIF test gave a sensitivity of 83.9% (95% CI: 66.3-94.5) and a specificity of 92.1% (95% CI: 83.6-97), a positive predictive value (PPV) of 81.3% (95% CI: 63.6-92.8) and a negative predictive value (NPV) of 93.3% (95% CI: 85.1-97.8). The GeneXpert MTB/RIF test had a fair sensitivity and specificity for diagnosing smear negative pulmonary TB. It may be useful for diagnosing pulmonary TB in patients with a negative sputum AFB smear. The assay is faster than culture and can detect rifampicin resistant strains of MTB.

RISK FACTORS FOR LATENT TUBERCULOSIS INFECTION AMONG HEALTH-CARE WORKERS IN NORTHEASTERN THAILAND.

Health-care workers (HCWs) are a high-risk population for acquiring Mycobacterium tuberculosis infection. Understanding the risk factors for latent tuberculosis infection (LTBI) could provide information to facilitate an appropriate tuberculosis (TB) control program. We aimed to determine the prevalence of, and risk factors for LTBI among HCWs in northeastern Thailand. Between 1 November 2013 and 30 September 2015, we examined 112 HCWs at Srinagarind Hospital, Khon Kaen Province in northeastern Thailand using the QuantiFERON®-TB Gold In-Tube (QFT) assay. Twenty-one [18.8%; 95% confidence interval (CI): 11.5- 26.0%] HCWs had a positive QFT result ­ all of whom were determined to have LTBI. The exposure risks and demographic data obtained from a questionnaire were compared between the 21 subjects who had a positive QFT assay and the 91 subjects who had a negative QFT assay. Multivariate analysis showed factors significantly associated with a positive QFT assay were: age ≥30 years (OR=18.88; 95%CI: 1.52-234.36), having worked as a nurse (OR=2.78; 95%CI: 1.19-6.49), having been employed at that job for ≥10 years (OR=8.78; 95%CI: 1.26-61.29) and having been exposed to known TB patients (OR=13.32: 95%CI: 1.61-110.04). Appropriate guidelines need to be developed, especially for these at-risk workers to prevent LTBI. These high-risk workers should also be considered for regular TB screening.

NOCARDIOSIS REVEALED BY THYROID ABSCESS AND PNEUMONIA IN A LIVER TRANSPLANT RECIPIENT.

Nocardia thyroid abscess with pneumonia is a rare clinical presentation. We reported a liver transplant recipient with Nocardia thyroiditis and pneumonia after receiving high dose immunosuppressants to preserve his graft. The patient is a 50-year-old male who developed hepatitis C virus-related liver cirrhosis and received a liver transplant. Seven months post-transplantation the patient developed graft rejection, which was treated with 3 days pulse dose methyl-prednisolone followed by an increased dose of his tracolimus, mycophenolate and prednisolone. He presented to the hospital with a 2 week history of fever, tenderness in his anterior neck and dry cough. On admission his temperature was 39.5°C. The right wing of his thyroid gland was swollen to 3 cm in size, fluctuant and tender. On auscultation of his lungs there were fine crepitations and increased vocal resonance in the right middle lung field. On laboratory testing, a complete blood count (CBC) revealed leukocytosis (19,900/mm3) with neutrophils (97%). A chest X-ray showed an patchy infiltrates and round circumscribed densities in the superior segment of the right lower lobe of his lung. A CT scan of his neck revealed a diffusely enlarged right wing of the thyroid gland, 3.8 cm in diameter that had an abnormal hyposignal area. A CT of his chest revealed consolidation of the superior segment of the right lower lobe and necrotic right paratracheal lymph nodes with inflamed strap muscles. Fine needle aspiration of the right lobe of thyroid gland was performed. Modified acid-fast bacilli (MAFB) staining showed partially acid-fast beaded branching filamentous organisms and a culture grew out Nocardia asteroides. He was treated with trimethoprim-sulfamethoxazole for 6 months. He improved clinically and his chest X-ray also cleared.

TRANSMISSION AND RISK FACTORS FOR LATENT TUBERCULOSIS INFECTIONS AMONG INDEX CASE-MATCHED HOUSEHOLD CONTACTS.

An understanding of the risk factors associated with acquiring and transmitting Mycobacterium tuberculosis (MTB) is required for controlling tuberculosis (TB). We aimed to determine the risk factors and transmission factors for latent tuberculosis infection (LTBI) in northeastern Thailand. Household contact persons (n = 70) and matched index patients with pulmonary TB (n = 42) who presented to Srinagarind Hospital, Khon Kaen, Thailand were interviewed from September 1, 2012 to March 31, 2014. LTBI was determined by positive results on both a tuberculin skin test and the QuantiFERON-TB Gold In-Tube test. Multivariate analysis of host and environmental risk factors was performed. Among contact persons, being aged 20 years (adjusted OR=14.0; 95% CI: 1.2-159.5), having a family relationship with a TB subject such as being a spouse or parent (adjusted OR=24.9; 95% CI: 2.4-263.9) and exposure to a TB subject for 5 hours/day (adjusted OR=9.2; 95% CI: 1.4-58.1) were risk factors for LTBI. Having a high bacillary load (adjusted OR=2; 95% CI: 1.26-3.17) or a moderate bacillary load (adjusted OR=1.39; 95% CI: 1.04-1.84) among TB subjects correlated with increased transmissibility compared to having a low bacillary load. The type of dwelling and density of household members were not found to be risk factors for LTBI in our study. We conclude being aged 20 years and having a relationship with a TB patient as a spouse or parent were risk factors for acquiring LTBI, and having a higher bacillary load was a risk factor for transmitting TB. Keywords: latent tuberculosis infection, transmission factor, risk factor, Mycobacterium tuberculosis, interferon-gamma release assay, Thailand

Evaluation of the QuantiFERON?-TB Gold In-Tube assay and tuberculin skin test for the diagnosis of Mycobacterium tuberculosis infection in northeastern Thailand.

BACKGROUND: The diagnosis of latent Mycobacterium tuberculosis infection (LTBI) is currently based on the immunological response of T-cells to M. tuberculosis (MTB) antigens. However, the QuantiFERON®-TB Gold In-Tube assay (QFT) has not yet been evaluated in the Thai adult population. OBJECTIVE: To evaluate the diagnostic performance and determine predictors of discordant results between the QFT and tuberculin skin test (TST). METHODS: Active tuberculosis (ATB) patients (n=54), close contacts (CCs) living in the same household as a TB patient (n=100) and healthy controls (HCs) (n=60) were interviewed and underwent the QFT and TST at Srinagarind Hospital in Thailand. Various cut-off values for the QFT (0.25-0.35 IU/mL) and TST (5-15 mm) were applied. RESULTS: The maximum agreement rate between the tests was 71.5% (κ=0.41) with cut-offs of 0.35 IU/mL and 10 mm or 0.25 IU/mL and 10 mm. Based on standard cut-off values (0.35 IU/mL and 10 mm) and using ATB patients and HCs as positive and negative controls, the TST was more sensitive than the QFT (87.0% vs. 66.7%, respectively), whereas the QFT was more specific than the TST (83.3% vs. 70.0%, respectively). Being underweight (OR 3.86, 95%CI 1.3-11.48) or overweight (OR 5.9, 95%CI 1.24-28.16) was significantly associated with TST+/QFT- results. Diabetes (OR 32.56, 95%CI 1.73-613.49) and poor or fair nutrition (OR 7.4, 95%CI 1.23-44.57) were significantly associated with TST-/QFT+ results. CONCLUSION: The TST should be used as a screening test based on its higher sensitivity, whereas the QFT should be used as a confirmatory test because of its higher specificity.

T-CELL RESPONSES ASSESSED USING IGRA AND TST ARE NOT CORRELATED WITH AFB GRADE AND CHEST RADIOGRAPH IN PULMONARY TUBERCULOSIS PATIENTS.

A definitive marker determining the bacillary load of Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB), and hence disease severity, is required for patient monitoring and management. In this study, the association among T-cell responses based on the interferon-gamma release assay (IGRA) and the tuberculin skin test (TST), the sputum acid-fast bacilli (AFB) grade and types of radiological lesions were analyzed in new cases of pulmonary TB patients (n = 54) at Srinagarind Hospital, Khon Kaen, Thailand between September 1, 2012 and March 31, 2014. It was found that infiltrative and cavitary lesions from chest radiographs were associated with high sputum AFB grade (p = 0.048). T-cell responses from both IGRA and TST were not correlated with sputum AFB grade. Neither IGRA nor TST was correlated with the bacillary load as defined by AFB grade and chest radiographs. Patients with cavitary lesions on chest radiographs tended to have high IFN-γ concentrations and large TST indurations. In addition, TB patients with previous BCG vaccination showed significantly higher IFN-γ induction compared to the non-vaccinated group (p = 0.001). This study showed T-cell responses based on both IGRA and TST were not correlated with AFB grade and chest radiograph. In areas of high rates of BCG vaccination, as in Thailand, the BCG may affect IGRA and TST interpretations.

Burden of adult pneumonia in Thailand: a nationwide hospital admission data 2010.

OBJECTIVE: To quantify the admission mortality, pathogens,factors related to mortality, length of hospital stay, and healthcare costs in adult hospitalized pneumonia in Thailand MATERIAL AND METHOD: The data on hospitalized pneumonia for the 2010 fiscal year extracted from the three main health insurance coverage schemes in Thailand (the Social Security System (SSS), the Medical Welfare System (MWS), and the Civil Servant Medical Benefit System (CSMBS)) were analyzed RESULTS: Adult hospitalized pneumonia admissions numbered 136,696, with mortality rate 9.63%. The mortality increased with increasing age, 15.49% for age > 80 years. Influenza virus was the major etiology for 19 to 25 years old (49.30%) with low mortality (1-2%). S. pneumoniae and typical pathogens were found in every age group. The mortality rate for S. pneumoniae increased with age, viz. 0%, 1.96%, 5.56%, 7.02%, 6.98%, and 24.24% for 19 to 25, 26 to 40, 41 to 60, 61 to 70, 71 to 80, and 81+ years old. The mortality rate from C. pneumonia was about 10% and high among the younger age group. Gram-negative bacilli and Staphylococcus caused high mortality (about 20 to 35%), especially in the older age group. The major risk factors for increasing mortality were: elderly (OR 3.46, 95% CI 3.27-3.77), alcoholic liver disease (OR 3.26, 95% CI 2.85-3.72), cirrhosis (OR 3.45, 95% CI 2.93-4.08), heart disease (OR 2.47, 95% CI 2.38-2.56), ischemic heart disease (OR 2.21, 95% CI 2.07-2.36), renal failure (OR 5.26, 95% CI 5.07-5.49), and cerebrovascular disease (OR 3.62, 95% CI 3.43-3.82). The median length of hospital stay was four days (IQR, 3-7 days) and the median cost of treatment per admission was US$ 256.63 (IQR, US$ 147.81-531.21). Complications such as acute respiratory failure, acute respiratory distress syndrome (ARDS), septicemia, shock, and acute renal failure made hospital stays two to three days longer and costs three to seven times higher than no complications. CONCLUSION: The mortality from pneumonia among the elderly was high, especially for those over 80 and with multiple medical co-morbidities.

Treatment outcomes of new and previously-treated smear positive pulmonary tuberculosis at Srinagarind Hospital, a tertiary care center in northeast Thailand.

OBJECTIVE: To study treatment outcomes of new and previously-treated smear positive pulmonary TB according to current WHO guideline 2010 and factors related to treatment success at Srinagarind Hospital. MATERIAL AND METHOD: Adult patients who had smear-positive pulmonary TB treated at Srinagarind Hospital between January 2005 and December 2010 were enrolled in the present study. RESULTS: Over a 6-years period, 322 patients (272 new and 50 previously-treated cases) were diagnosed smear positive pulmonary TB. The mean age was 48.85 (SD 17.9) years and the male to female ratio was 1.8:1. The mean duration of symptoms in the previously-treated group was longer than the new cases (2.39 vs. 1.99 months, p = 0.38). Symptoms, underlying diseases, HIV status, and organ involvement between these two groups were not different. Two-thirds (72.1% of patients) had cough, 35.4% had fever and 20.5% weight loss. Twenty-three percent of cases had underlying DM and 7.8% were HIV positive. Disseminated TB was found in 18.9% of cases. The mean duration of treatment in new cases was 6.88 months vs. 11.20 months in previously-treated cases. The common regimens for new cases included 2IRZE/4IR (72.8%) and 2IRE/7IR (19.1%) vs. 2IRZE/4IR (62%), 2IRE/7IR (12%), and other regimens for MDR patients (6%) among the previously-treated cases. However in previously-treated group IRZES/IRZE/IRE (p = 0.001), second-line drugs (p = 0.002), and MDR regimens (p < 0.001) were statistically more common treatments than in the new cases group. About 60% of cases were treated at TB clinic. The success rate among new cases who had completed treatment at Srinagarind Hospital was higher than for previously-treated ones (94.8% vs. 86.4%; p = 0.04). Among previously-treated cases (n = 50); 24 were defaulters, 19 were relapses, and seven were failures. For the defaulted and relapsed cases, patients usually received the IRZE/IR or IRZES/IRZE/IRE regimens. On the other hand, for failure cases, patients usually received the second-line drugs or MDR regimens. The overall success rate in defaulted cases was 87.6%, vs. 68.4% who relapsed and 57.1% who failed (p = 0.067). For new cases, the isolations found DR-TB 3.6% and MDR-TB 0%. For previously-treated cases, the isolations found DR-TB 16.67% and MDR-TB 6.25%. The only one factor related to successful outcomes was treatment at TB clinic (adjusted OR 2.01, 95% CI 1.18-3.43). CONCLUSION: Previously-treated pulmonary TB had less success rate than new cases. Culture and susceptibility for previously-treated group were recommended before starting treatment. Treatment at TB clinic improved treatment outcomes.

Determination of latent tuberculosis infection from plasma samples via label-free SERS sensors and machine learning.

Effective diagnostic tools for screening of latent tuberculosis infection (LTBI) are lacking. We aim to investigate the performance of LTBI diagnostic approaches using label-free surface-enhanced Raman spectroscopy (SERS). We used 1000 plasma samples from Northeast Thailand. Fifty percent of the samples had tested positive in the interferon-gamma release assay (IGRA) and 50 % negative. The SERS investigations were performed on individually prepared protein specimens using the Raman-mapping technique over a 7 × 7 grid area under measurement conditions that took under 10 min to complete. The machine-learning analysis approaches were optimized for the best diagnostic performance. We found that the SERS sensors provide 81 % accuracy according to train-test split analysis and 75 % for LOOCV analysis from all samples, regardless of the batch-to-batch variation of the sample sets and SERS chip. The accuracy increased to 93 % when the logistic regression model was used to analyze the last three batches of samples, following optimization of the sample collection, SERS chips, and database. We demonstrated that SERS analysis with machine learning is a potential diagnostic tool for LTBI screening.

Causative agents and resistance among hospital-acquired and ventilator-associated pneumonia patients at Srinagarind Hospital, northeastern Thailand.

Abstract. Hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP) have an impact on health care costs and mortality. The aim of this study was to identify the causative agents, antibiotics prescribed, cost of treatment and drug resistance trends among HAP and VAP patients at a tertiary-care hospital in northeastern Thailand during 2008 and 2009. The incidences of HAP in 2008 and 2009 were 0.7/1,000 and 0.55/1,000 hospital days, respectively. The incidences of VAP in 2008 and 2009 were 13.6/1,000 and 12.6/1,000 ventilator days, respectively. About 70% of HAP were caused by Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae; and 70% of VAP were caused by A. baumannii, P. aeruginosa, and K. pneumoniae. The ranking in the causative agents of HAP and VAP was not different, but more antimicrobial resistant organisms were seen in 2009. More than half of the costs of nosocomial infection treatment in 2008 and 2009 were the costs for HAP and VAP, 16.8 and 17.5 million Baht, respectively. Fewer A. baumannii and P. aeruginosa isolates were sensitive to carbapenems. Only one-fifth of A. baumannii isolates were sensitive to cefoperazone/sulbactam. The only two antimicrobial agents with consistently good activity against A. baumannii were tigecycline (approximately 85%) and colistin (approximately 99%). Fifty-seven point six percent of P. aeruginosa isolates were sensitive to ceftazidime, 72.4% were sensitive to piperacillin/tazobactam, 95.9% were sensitive to netilmycin and 99.2% were sensitive to colistin. Forty-seven percent of K. pneumoniae isolates were extended spectrum beta-lactamase and sensitive to carbapenems. Methicillin-resistant S. aureus was the cause of 6-7% of HAP/VAP cases in our study.

Serum proteomics analysis for differentiation among Mycobacterium tuberculosis infection categories.

Inhalation of Mycobacterium tuberculosis (Mtb) bacilli can lead to a range of TB categories including early clearance (EC), latent TB infection (LTBI) and active TB (ATB). There are few biomarkers available to differentiate among these TB categories: effective new biomarkers are badly needed. Here, we analyzed the serum proteins from 26 ATB cases, 20 LTBI cases, 34 EC cases and 38 healthy controls (HC) using label-free LC-MS/MS. The results were analyzed using MaxQuant software and matched to three different bacterial proteomics databases, including Mtb, Mycobacterium spp. and normal lung flora. PCA of protein candidates using the three proteomics databases revealed 44.5% differentiation power to differentiate among four TB categories. There were 289 proteins that showed potential for distinguishing between each pair of groups among TB categories. There were 50 candidate protein markers specifically found in ATB and LTBI but not in HC and EC groups. Decision trees using the top five candidate biomarkers (A0A1A2RWZ9, A0A1A3FMY8, A0A1A3KIY2, A0A5C7MJH5 and A0A1X0XYR3) had 92.31% accuracy to differentiate among TB categories and the accuracy was increased to 100% when using 10 candidate biomarkers. Our study shows that proteins expressed from Mycobacterium spp. have the potential to be used to differentiate among TB categories.

High clustering rate and genotypic drug-susceptibility screening for the newly recommended anti-tuberculosis drugs among global extensively drug-resistant Mycobacterium tuberculosis isolates.

Multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) make TB difficult to control. Global susceptibility data for six newly recommended anti-TB drugs against M/XDR-TB are still limited. Using publicly available whole-genome sequences, we determined the proportion of 513 phenotypically XDR-TB isolates that carried mutations associated with resistance against these drugs (bedaquiline, clofazimine, linezolid, delamanid, pretomanid and cycloserine). Mutations of Rv0678 and Rv1979c were detected in 69/513 isolates (13.5%) for bedaquiline resistance and 79/513 isolates (15.4%) for clofazimine resistance with additional mmpL5 mutations. Mutations conferring resistance to delamanid were detected in fbiB and ddn genes for 11/513 isolates (2.1%). For pretomanid, a mutation was detected in the ddn gene for 3/513 isolates (0.6%). Nineteen mutations of pykA, cycA, ald, and alr genes, conferring resistance to cycloserine, were found in 153/513 isolates (29.8%). No known mutations associated with linezolid resistance were detected. Cluster analysis showed that 408/513 isolates fell within 99 clusters and that 354 of these isolates were possible primary drug-resistant TB (292 XDR-TB, 57 pre-XDR-TB and 5 MDR-TB). Clonal transmission of primary XDR isolates might contribute significantly to the high prevalence of DR-TB globally.

Drug susceptibility patterns of Mycobacterium tuberculosis and clinical outcomes of drug-resistant tuberculosis at Srinagarind Hospital, a tertiary care center in northeastern Thailand.

Drug-resistant tuberculosis is a major public health problem. The aim of this study was to assess the local susceptibility patterns of Mycobacterium tuberculosis and clinical outcomes of drug resistant tuberculosis (DR-TB) at Srinagarind Hospital, a tertiary care center in northeastern Thailand. Between January 2004 and December 2008, 1,052 patients had culture-proven M. tuberculosis infections at Srinagarind Hospital. M. tuberculosis was resistant to isoniazid (2.3%), rifampicin (2.8%), ethambutol (3.8%), streptomycin (2.1%), kanamycin (0.7%) and ofloxacin (1.9%). The occurrences of multi-drug resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB) were 1.2% and 0.38%, respectively. Of the 65 DR-TB patients, complete medical records were found for 55. The male to female ratio was 2.2:1. The mean age was 50 years. Thirteen patients had MDR-TB. The duration of symptoms in the MDR-TB group was longer than the non-MDR-TB group, 11.6 months vs 2.6 months, respectively. Half of MDR-TB and one-third of non-MDR-TB patients had a previous history of being treated for tuberculosis. Nearly 20% of cases were HIV positive. Mono-drug resistance was initially treated with standard first-line drugs (CAT 1). The clinical course was more likely to be worse during the maintenance phase if there was resistance to rifampicin. Whenever there was resistance to two, three or four drugs, the antituberculosis drugs were prescribed based on susceptibility patterns. Only 30% of patients with MDR-TB and XDR-TB responded to treatment. Culture and sensitivity testing for M. tuberculosis cases is recommended in patients at high risk for DR-TB, such as patients previously treated for tuberculosis and those HIV positive.

Presentations of chronic cavitary pulmonary histoplasmosis mimic infected cystic bronchiectasis in an immunocompetent host: A case report.

BACKGROUND: Chronic cavitary pulmonary disease and laryngeal involvement are unusual manifestations of Histoplasmosis capsulatum infection, particularly in patients who are not immunocompromised. The presence of fibro-cavitary lesions has been reported as a radiologic presentation of chronic histoplasmosis in patients with pre-existing lung disease. However, there have been few reports of extensive basal predominant cavitary lesions that mimic cystic-bronchiectasis. CASE PRESENTATION: A 65-year-old previously healthy Thai male presented with productive cough, hoarseness, low-grade fever, and weight loss for 6 months. There was no history of significant exposure to Histoplasmosis capsulatum. Tests for HIV and anti-IFN- γ antibody were negative. Chest CT revealed multifocal thick wall cavities, which were distributed in a peri-bronchial pattern, and some areas of consolidation in both basal lungs. Laryngoscopy revealed an ulcerative lesion of the false vocal cords. Histopathological study of false vocal cords and lung tissue showed granulomatous inflammation with mixed inflammatory cell infiltration and aggregation of histiocytes containing round intracytoplasmic organisms. GMS-staining was positive, but negative mucicarmine-staining was negative. A real-time PCR assay of the lung tissue was positive for Histoplasmosis capsulatum. The final diagnosis was chronic cavitary pulmonary histoplasmosis with laryngeal involvement. CONCLUSION: Chronic cavitary pulmonary histoplasmosis is rare, as is laryngeal involvement. However, there have been such cases in endemic areas, even in immunocompetent patients. Chronic histoplasmosis should be considered in patients who present with the extensive basal predominant cavitary-pulmonary lesions that mimic cystic bronchiectasis.

Whole-genome analysis of drug-resistant Mycobacterium tuberculosis reveals novel mutations associated with fluoroquinolone resistance.

Multidrug-resistant and extensively drug-resistant tuberculosis (M/XDR-TB) remains a global public-health challenge. Known mutations in quinolone resistance-determination regions cannot fully explain phenotypic fluoroquinolone (FQ) resistance in Mycobacterium tuberculosis (Mtb). The aim of this study was to look for novel mutations in Mtb associated with resistance to FQ drugs using whole-genome sequencing analysis. Whole-genome sequences of 659 Mtb strains, including 214 with phenotypic FQ resistance and 445 pan-susceptible isolates, were explored for mutations associated with FQ resistance overall and with resistance to individual FQ drugs (ofloxacin, levofloxacin, moxifloxacin and gatifloxacin). Three novel genes (recC, Rv2005c and PPE59) associated with FQ resistance were identified (P < 0.00001 based on screening analysis and absence of relevant mutations in a pan-susceptible validation set of 360 strains). Nine novel single nucleotide polymorphisms (SNPs), including in gyrB (G5383A and G6773A), gyrA (G7892A), recC (G725900C and G726857T/C), Rv2005c (C2251373G, G2251420C and C2251725T) and PPE59 (C3847269T), were used for diagnostic performance analysis. Enhancing the known SNP set with five of these novel SNPs, including gyrA [G7892A (Leu247Leu)], recC [G725900C (Leu893Leu) and G726857T/C (Arg484Arg)], Rv2005c [G2251420C (Pro205Arg)] and PPE59 [C3847269T (Asn35Asn)] increased the sensitivity of detection of FQ-resistant Mtb from 83.2% (178/214) to 86.9% (186/214) while maintaining 100% specificity (360/360). No specific mutation associated with resistance to only a single drug (ofloxacin, levofloxacin, moxifloxacin or gatifloxacin) was found. In conclusion, this study reports possible additional mutations associated with FQ resistance in Mtb.