RESUMO
PURPOSE: To investigate urine microbiome differences among healthy women, women with recurrent uncomplicated cystitis (rUC), and those with sporadic/single uncomplicated cystitis (sUC) to challenge traditional beliefs about origins of these infections. MATERIALS AND METHODS: Patients who underwent both conventional urine culture and next-generation sequencing (NGS) of urine were retrospectively reviewed. Symptom-free women with normal urinalysis results as a control group were also studied. Samples were collected via transurethral catheterization. RESULTS: In the control group, urine microbiome was detected on NGS in 83.3%, with Lactobacillus and Prevotella being the most abundant genera. The sensitivity of urine NGS was significantly higher than that of conventional urine culture in both the sUC group (91.2% vs. 32.4%) and the rUC group (82.4% vs. 16.4%). In urine NGS results, Enterobacterales, Prevotella, and Escherichia/Shigella were additionally found in the sUC group, while the recurrent urinary tract infection (rUTI)/rUC group exhibited the presence of Lactobacillus, Prevotella, Enterobacterales, Escherichia/Shigella, and Propionibacterium. Moreover, distinct patterns of urine NGS were observed based on menopausal status and ingestion of antibiotics or probiotics prior to NGS test sampling. CONCLUSIONS: Urine microbiomes in control, sUC, and rUTI/rUC groups exhibited distinct characteristics. Notably, sUC and rUC might represent entirely separate pathological processes, given their distinct urine microbiomes. Consequently, the use of urine NGS might be essential to enhancing sensitivity compared to conventional urine culture in both sUC and rUTI/rUC groups.
Assuntos
Cistite , Microbiota , Recidiva , Humanos , Feminino , Cistite/microbiologia , Cistite/urina , Estudos Retrospectivos , Pessoa de Meia-Idade , Adulto , Urina/microbiologia , República da Coreia , Sequenciamento de Nucleotídeos em Larga Escala , Doença Aguda , Infecções Urinárias/microbiologia , Idoso , Relevância ClínicaRESUMO
In this observational study conducted in 2022, 12.3% of patients who shared a room with a patient positive for severe acute respiratory coronavirus virus 2 (SARS-CoV-2) also had a positive polymerase chain reaction (PCR) test, either at initial screening or during a 5-day quarantine. Therefore, screening and quarantine are still necessary within hospitals for close-contact inpatients during the SARS-CoV-2 omicron-variant dominant period.
Assuntos
COVID-19 , Viroses , Humanos , SARS-CoV-2/genética , Pacientes Internados , COVID-19/diagnóstico , Reação em Cadeia da Polimerase , Teste para COVID-19RESUMO
Accurate detection and classification of white blood cells, otherwise known as leukocytes, play a critical role in diagnosing and monitoring various illnesses. However, conventional methods, such as manual classification by trained professionals, must be revised in terms of accuracy, efficiency, and potential bias. Moreover, applying deep learning techniques to detect and classify white blood cells using microscopic images is challenging owing to limited data, resolution noise, irregular shapes, and varying colors from different sources. This study presents a novel approach integrating object detection and classification for numerous type-white blood cell. We designed a 2-way approach to use two types of images: WBC and nucleus. YOLO (fast object detection) and ViT (powerful image representation capabilities) are effectively integrated into 16 classes. The proposed model demonstrates an exceptional 96.449% accuracy rate in classification.
Assuntos
Interpretação de Imagem Assistida por Computador , Leucócitos , Aprendizado Profundo , MicroscopiaRESUMO
A 65-year-old male patient with an end-stage renal disease was diagnosed with coronavirus disease 2019 (COVID-19) by reverse transcription polymerase chain reaction. The patient complained of cough, sputum, and respiratory distress that worsened three days ago. The patient required mechanical ventilation and extracorporeal mentrane oxygenation. On day 9, convalescent plasma collected from a 34-year old man who recovered from COVID-19 45 days ago was administered. The patient showed immediate clinical improvement. However, on day 14, the patient's clinical course worsened again. On day 19 and day 24, vancomycin-resistant Enterococcus faecium bacteremia and methicillin-resistant Staphylococcus aureus pneumonia were found. After long-term supportive care, he slowly recovered. He was discharged on day 91 without any oxygen requirement. This case report suggests that convalescent plasma therapy might just provide a short-term relief and that persistent effort for critical care is necessary to save patients from severe COVID-19.
RESUMO
Recent studies on the urine microbiome have highlighted the importance of the gut-vagina-bladder axis in recurrent urinary tract infection (rUTI). In particular, the role of Gardnerella as a covert pathogen that activates E. coli in animal experiments has been reported. Herein, we conducted a human bladder microbiome study to investigate the effect of Gardnerella on rUTI. Urine 16S ribosomal RNA gene sequencing via transurethral catheterization was conducted in the normal control group (NC) (n = 18) and rUTI group (n = 78). The positive detection rate of Gardnerella species did not differ between the NC and rUTI groups (22.2% vs. 18.0%, p = 0.677). In addition, the Gardnerella-positive NC and Gardnerella-positive rUTI groups showed similar levels of microbiome diversity. The Gardnerella-positive group was categorized into three subgroups: the Escherichia-dominant group, Gardnerella-dominant group, and Lactobacillus-dominant group. All of the Escherichia-dominant groups were associated with rUTI. The Gardnerella-dominant or Lactobacillus-dominant groups expressed rUTI with symptoms when risk factors such as the degree of Gardnerella proliferation or causative agents of bacterial vaginosis were present. The presence of Gardnerella in the urine is considered to be related to rUTI depending on other risk factors. New guideline recommendations regarding antibiotic selection based on a novel method to detect the cause of rUTI may be required to reduce antibiotic resistance.
RESUMO
Traditionally, the diagnostic mainstay of recurrent urinary tract infection has been urinary culture. However, the causative uropathogen of recurrent cystitis has not been well established. Urine DNA next-generation sequencing (NGS) can provide additional information on these infections. Herein, we compared urine NGS results and urine cultures in patients with acute uncomplicated cystitis (AUC) and recurrent cystitis (RC), and evaluated the difference in microbiome patterns in the NGS results. Patients who underwent urine culture and NGS due to AUC or RC were retrospectively reviewed. All urine samples were collected via a transurethral catheter and studied utilizing a type of NGS called 16S ribosomal RNA gene amplification and sequencing. The sensitivity of urine NGS was significantly higher than that of conventional urine culture (69.0% vs. 16.7%, p < 0.05). The detection rate of urine NGS was slightly lower in the RC group than in the AUC group (67.7% vs. 72.7%). Microbiome diversity was significantly higher in the RC group compared to the AUC group (p = 0.007), and the microbiome composition was significantly different between the AUC and RC groups. In the urine NGS results, Pseudomonas, Acinetobacter, and Enterobacteriaceae were found in the AUC group, and Sphingomonas, Staphylococcus, Streptococcus, and Rothia spp. were detected in the RC group. Urine NGS can significantly increase the diagnostic sensitivity compared to traditional urine culture methods, especially in RC patients. AUC and RC patients had significant differences in bacterial diversity and patterns. Therefore, recurrent cystitis might be approached from a different perspective.
RESUMO
Cytogenetic dosimetry is useful for evaluating the absorbed dose of ionizing radiation based on analysis of radiation-induced chromosomal aberrations. We created two types of in vitro dose-response calibration curves for dicentric chromosomes (DC) and translocations (TR) induced by X-ray irradiation, using an electron linear accelerator, which is the most frequently used medical device in radiotherapy. We irradiated samples from four healthy Korean individuals and compared the resultant curves between individuals. Aberration yields were studied in a total of 31,800 and 31,725 metaphases for DC and TR, respectively, obtained from 11 X-ray irradiation dose-points (0, 0.05, 0.1, 0.25, 0.5, 0.75, 1, 2, 3, 4, and 5 Gy). The dose-response relationship followed a linear-quadratic equation, Y=C+αD+ßD², with the coefficients C=0.0011 for DC and 0.0015 for TR, α=0.0119 for DC and 0.0048 for TR, and ß=0.0617 for DC and 0.0237 for TR. Correlation coefficients between irradiation doses and chromosomal aberrations were 0.971 for DC and 0.6 for TR, indicating a very strong and a moderate correlation, respectively. This is the first study implementing cytogenetic dosimetry following exposure to ionizing X-radiation.
Assuntos
Linfócitos/efeitos da radiação , Radiação Ionizante , Adulto , Povo Asiático , Aberrações Cromossômicas/efeitos da radiação , Feminino , Humanos , Cariotipagem , Linfócitos/metabolismo , Masculino , Radiometria , República da Coreia , Translocação Genética/efeitos da radiação , Adulto JovemRESUMO
Human epididymis protein 4 (HE4) has been suggested as a useful new biomarker of lung cancer; however, few relevant large-scale studies have been published. In this study, we evaluated the utility of serum HE4 for lung cancer detection. HE4 levels were measured in serum samples from 100 lung cancer patients, 57 patients with benign lung diseases, and 274 healthy controls by using a chemiluminescent immunoassay, and variations in HE4 levels were analyzed by clinical status such as lung cancer, benign lung disease, and healthy condition, Tumor, Lymph Nodes, Metastasis (TNM) stage, tumor score, and histological cancer type. Lung cancer patients had significantly higher serum HE4 levels than patients with benign lung diseases and healthy controls (P<0.0001). The area under the ROC curve for HE4 was 0.84 (95% confidence interval, 0.78-0.89; P<0.0001) between lung cancer patients and healthy controls. Serum HE4 levels were significantly higher in patients with advanced disease (according to TNM stage) than in healthy controls (P<0.0001). HE4 levels were significantly elevated in patients with tumors of all types, those of different histological subgroups, and those with the smallest tumors (P=0.002). This report supports the potential of serum HE4 as an ancillary diagnostic marker for lung cancer detection.
Assuntos
Biomarcadores Tumorais/sangue , Imunoensaio , Neoplasias Pulmonares/diagnóstico , Proteínas/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Estudos de Casos e Controles , Feminino , Humanos , Medições Luminescentes , Neoplasias Pulmonares/patologia , Linfonodos/patologia , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Estadiamento de Neoplasias , Curva ROC , Proteína 2 do Domínio Central WAP de Quatro DissulfetosRESUMO
OBJECTIVES: To identify bacterial contamination rates of laryngoscope blades and handles stored in emergency crash carts by hospital and area according to the frequency of intubation attempts. METHODS: One hundred forty-eight handles and 71 blades deemed ready for patient use from two tertiary hospitals were sampled with sterile swabs using a standardized rolling technique. Samples were considered negative (not contaminated) if no colonies were present on the blood agar plate after an 18-hour incubation period. Samples were stratified by hospital and according to the frequency of intubation attempts (10 attempts per year) using the χ2-test and Fisher exact test. RESULTS: One or more species of bacteria were isolated from 4 (5.6%) handle tops, 20 (28.2%) handles with knurled surfaces, and 27 (18.2%) blades. No significant differences were found in microbial contamination levels on the handle tops and blades between the two hospitals and two areas according to the frequency of intubation attempts. However, significant differences were found between the two hospitals and two areas in the level of microbial contamination on the handles with knurled surfaces (p<0.05). CONCLUSIONS: Protocols and policies must be reviewed to standardize procedures to clean and disinfect laryngoscope blades and handles; handles should be re-designed to eliminate points of contact with the blade; and single-use, one-piece laryngoscopes should be introduced.
Assuntos
Desinfecção/métodos , Equipe de Respostas Rápidas de Hospitais , Intubação Intratraqueal/instrumentação , Laringoscópios/microbiologia , Infecção Hospitalar/prevenção & controle , Humanos , Estudos Prospectivos , República da Coreia , Centros de Atenção TerciáriaAssuntos
Subunidade alfa 2 de Fator de Ligação ao Core/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Proteínas de Fusão Oncogênica/genética , Medula Óssea/patologia , Cromossomos Humanos Par 12 , Cromossomos Humanos Par 9 , DNA/metabolismo , Rearranjo Gênico , Humanos , Hibridização in Situ Fluorescente , Cariotipagem , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Translocação GenéticaRESUMO
BACKGROUND: Extensively drug-resistant (XDR) Pseudomonas aeruginosa and Acinetobacter baumannii are a threat to hospitalized patients. We evaluated the effects of antimicrobial combinations on XDR P. aeruginosa and A. baumannii isolates. METHODS: P. aeruginosa and A. baumannii isolates, which were resistant to all antibiotics except colistin (CL), were collected from eight hospitals in Korea. Genes encoding metallo-ß-lactamases (MBLs) and OXA carbapenemases were detected by PCR in eight P. aeruginosa and 30 A. baumannii isolates. In vitro synergy of antimicrobial combinations was tested by using the checkerboard method. RESULTS: Minimum inhibitory concentrations of ß-lactams, aminoglycosides, and fluoroquinolones were very high, while that of CL was low for majority of XDR P. aeruginosa and A. baumannii isolates. Antimicrobial combinations including Imipenem (IPM)-CL, ceftazidime (CAZ)-CL, and rifampin (RIF)-CL exerted only additive/indifferent effects on majority of XDR P. aeruginosa isolates. Proportions of XDR A. baumannii isolates that showed synergistic and additive/indifferent inhibition after treatment with antimicrobial combinations used are as follows: IPM-ampicillin-sulbactam (AMS), 17% and 80% isolates, respectively; IPM-rifampin (RIF), 13% and 81% isolates, respectively; IPM-CL, 13% and 87% isolates, respectively; and RIF-COL, 20% and 73% isolates, respectively. Significant proportion (19%) of XDR P. aeruginosa isolates produced MBLs, and majority (82%) of A. baumannii isolates produced either MBLs or OXA-23. CONCLUSIONS: Our results suggest that combinations of IPM-AMS, IPM-RIF, IPM-CL, and RIF-CL are more useful than individual drugs for treating 13-20% of XDR A. baumannii infections.
Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Anti-Infecciosos/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Acinetobacter baumannii/genética , Acinetobacter baumannii/isolamento & purificação , Aminoglicosídeos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sinergismo Farmacológico , Fluoroquinolonas/farmacologia , Imipenem/farmacologia , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , beta-Lactamases/genética , beta-Lactamases/metabolismoRESUMO
PURPOSE: The genus Aeromonas is a pathogen that is well known to cause severe clinical illnesses, ranging from gastroenteritis to sepsis. Accurate identification of A. hydrophila, A. caviae, and A. veronii is important for the care of patients. However, species identification remains difficult using conventional methods. The aim of this study was to compare the accuracy of different methods of identifying Aeromonas at the species level: a biochemical method, matrix-assisted laser desorption ionization mass spectrometry-time of flight (MALDI-TOF MS), 16S rRNA sequencing, and housekeeping gene sequencing (gyrB, rpoB). MATERIALS AND METHODS: We analyzed 65 Aeromonas isolates recovered from patients at a university hospital in Korea between 1996 and 2012. The isolates were recovered from frozen states and tested using the following four methods: a conventional biochemical method, 16S rRNA sequencing, housekeeping gene sequencing with phylogenetic analysis, and MALDI-TOF MS. RESULTS: The conventional biochemical method and 16S rRNA sequencing identified Aeromonas at the genus level very accurately, although species level identification was unsatisfactory. MALDI-TOF MS system correctly identified 60 (92.3%) isolates at the species level and an additional four (6.2%) at the genus level. Overall, housekeeping gene sequencing with phylogenetic analysis was found to be the most accurate in identifying Aeromonas at the species level. CONCLUSION: The most accurate method of identification of Aeromonas to species level is by housekeeping gene sequencing, although high cost and technical difficulty hinder its usage in clinical settings. An easy-to-use identification method is needed for clinical laboratories, for which MALDI-TOF MS could be a strong candidate.
Assuntos
Aeromonas/genética , Genes Essenciais/genética , Tipagem Molecular/métodos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Aeromonas/classificação , Aeromonas/isolamento & purificação , DNA Bacteriano/genética , Humanos , Filogenia , República da Coreia , Sensibilidade e EspecificidadeAssuntos
Malária Vivax/diagnóstico , Neutrófilos/parasitologia , Plasmodium vivax/isolamento & purificação , Adulto , Antibacterianos/uso terapêutico , Antimaláricos/uso terapêutico , Clindamicina/uso terapêutico , Feminino , Humanos , Malária Vivax/tratamento farmacológico , Malária Vivax/parasitologia , Plasmodium vivax/crescimento & desenvolvimento , Gravidez , Quinina/uso terapêutico , Trofozoítos/citologiaRESUMO
BACKGROUND: Antimicrobial surveillance is important for providing an up-to-date understanding of the epidemiology of antimicrobial resistance and for creating a forum for rational drug development. In this study, we analyzed antimicrobial test data generated in 2011 by hospitals and commercial laboratories participating in the Korean Nationwide Surveillance of Antimicrobial Resistance program (KONSAR). MATERIALS AND METHODS: Data on the results of susceptibility tests conducted in 32 hospitals and two commercial laboratories were analyzed. Data on isolates from patients admitted to an intensive care unit (ICU) and those admitted to other wards were compared. Intermediate susceptibility was not analyzed and duplicate isolates were excluded. RESULTS: Escherichia coli was the most prevalent organism identified in both the hospital and commercial laboratories. Among the hospital isolates, methicillin-resistant Staphylococcus aureus (MRSA), penicillin G-non-susceptible Streptococcus pneumoniae, and ampicillin-resistant Enterococcus faecium remained as prevalent as they were in 2009. The proportion of vancomycin-resistant E. faecium (VR-EFM) slightly decreased from 29% in 2009 to 23% in 2011. Resistance rates of Klebsiella pneumoniae to ceftazidime, cefoxitin, fluoroquinolone, and amikacin were 24%, 14%, 27%, and 8%, respectively. Resistance rates of Pseudomonas aeruginosa to fluoroquinolone, ceftazidime, imipenem, and amikacin were 33%, 20%, 22%, and 16%, respectively, whereas those of Acinetobacter spp. resistance were 71%, 66%, 64, and 51%, respectively. The prevalence of oxyimino-cephalosporin-resistant E. coli and K. pneumoniae, carbapenem-resistant Acinetobacter spp. and P. aeruginosa, MRSA, and VR-EFM among ICU isolates was higher than those among non-ICU isolates. Extended-spectrum ß-lactamase-producing E. coli and K. pneumoniae, imipenem-resistant P. aeruginosa, and VR-EFM were more prevalent among isolates from commercial laboratories than those from hospitals. Resistance rates of K. pneumoniae to ceftazidime and amikacin decreased from 32% and 24% in 2005 to 24% and 8% in 2011, respectively. The resistance rate of P. aeruginosa to amikacin decreased from 22% in 2005 to 16% in 2011. The proportion of imipenem-resistant Acinetobacter spp. increased from 16% in 2005 to 64% in 2011. CONCLUSIONS: The prevalence of MRSA, penicillin G-non-susceptible S. pneumoniae, and ampicillin-resistant E. faecium among clinical isolates tested in laboratories remained high. Multidrug resistance was more prevalent among isolates from ICUs. The prevalence of ceftazidime-resistant and amikacin-resistant K. pneumoniae and amikacin-resistant P. aeruginosa decreased after 2005, while the prevalence of imipenem-resistant Acinetobacter spp. increased.
RESUMO
Mycobacterium neoaurum is rapidly growing mycobacteria that can cause human infections. It commonly causes bloodstream infections in immunocompromised hosts, and unlike other mycobacteria species, it rarely causes pulmonary infections. We confirmed the first pulmonary infection case in Korea caused by M. neoaurum using full-length 16S rRNA gene sequencing.
Assuntos
Pneumopatias/diagnóstico , Infecções por Mycobacterium/diagnóstico , Mycobacterium/isolamento & purificação , Adulto , Feminino , Humanos , Pneumopatias/microbiologia , Mycobacterium/genética , Infecções por Mycobacterium/microbiologia , Micobactérias não Tuberculosas/genética , Micobactérias não Tuberculosas/isolamento & purificação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de RNARESUMO
UNLABELLED: CuraVAC Ag, a product that delivers negative pressure wound therapy through a polyurethane foam dressing, contains silver nanoparticles, which, when moistened with water, release silver ions onto a wound surface. The in vitro antimicrobial action of silver can destroy both gram-positive and gram-negative bacteria, as well as methicillin-resistant Staphylococcus aureus (MRSA). The purpose of this study was to assess the efficacy and in vivo outcomes of using the product. METHODS: Thirty-six female Sprague-Dawley white rats, 8-weeks old and 250 g - 300 g in weight, were used. The experimental product was prepared using a vacuum-assisted closure (VAC) kit and coating it using the silver nanoparticles. For the control group, a 10% povidone-iodine solution was applied. RESULTS: All groups showed decreases in wound area over time, in the order CuraVAC Ag (group A) > CuraVAC (group B) > control (group C). On the third, fifth, and seventh days, wound healing efficacy scores increased in both group A and group C. Groups A and B showed more rapid decreases than group C in bacterial culture from wounds. CONCLUSION: CuraVAC Ag may be useful for treatment of wounds infected with bacteria..
RESUMO
OBJECTIVE: Silver plays an important part in severe wound management, mainly by reducing microbial growth within dressed wounds and wound beds. However, it is unknown how silver-coated dressing materials affect diabetic wounds. The purpose of this study is to evaluate the efficacy of silver-containing dressing materials for the treatment of methicillin-resistant Staphylococcus aureus (MRSA)-infected wounds in streptozotocininduced diabetic rats. METHODS: Full-thickness skin defects were created on the backs of rats with streptozotocin (STZ)-induced diabetes (n = 108) and were infected with MRSA. The rats were assigned to 6 groups according to the wound dressing used: nanocrystalline silver (Acticoat, Smith & Nephew, Inc, London, UK), silver carboxymethylcellulose (Aquacel-Ag, ConvaTec, Skillman, NJ), silver sulfadiazine (Medifoam silver, Genewel Science Co Ltd, Seongnam, South Korea), nanocrystalline silver (Poly- Mem silver, Ferris Mfg Corp, Burr Ridge, IL), silver sulfadiazine (Ilvadon, Ildong Pharmaceuticals, Seoul, South Korea), and 10% povidone iodide (control). The wound size, histological findings, and bacterial colony count for each group was analyzed and compared to normal Sprague-Dawley rats. RESULTS: Wound size decreased over time in every group. On day 10, a significant difference in wound area was detected between the silver dressing groups and the control group (P = 0.0040). In the wound biopsy, on days 4, 7, and 10, the would-healing effect increased in every group. However, between days 4 (P = 0.8250) and 10 (P = 0.9912), there was no statistical difference between groups. The number of bacteria in each group decreased with time in the bacterial wound culture. The silver dressing groups were more effective on antimicrobial efficacy, but there was no statistically significant difference between the silver dressing groups and the control group. CONCLUSION: Silver-containing dressing materials are an effective method for MRSA-infected wounds, but nano silver-containing dressing materials did not have better results in a diabetic rat model compared to a normal rat model in historical review. Further investigation is necessary to determine the relative safety of these products on the healing wound. Once that is done, the relative value of the products can be determined by balancing their antimicrobial and cytotoxicity characteristics.
RESUMO
Common bacterial pathogens of otitis media include Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, alpha-hemolytic streptococci, and Group A streptococci. We recently isolated a gram-negative, rod-shaped, non-spore-forming bacterium from a patient with otitis media following tympanocentesis. 16S rRNA gene sequence similarity studies of effusion identified this strain (CCUG 43427AT) as Massilia sp. (99.7%). Massilia spp. have been isolated from soil, air, and immunocompromised patients. However, there are no reports of their isolation from cases of otitis media. This case report highlights a rare and novel bacterial organism of otitis media.
Assuntos
Otite Média com Derrame/microbiologia , Oxalobacteraceae/isolamento & purificação , Criança , Humanos , Masculino , Otite Média com Derrame/diagnóstico , Otite Média com Derrame/cirurgia , Reação em Cadeia da PolimeraseRESUMO
BACKGROUND: A peptide nucleic acid (PNA) probe-based real-time PCR (PNAqPCR™ TB/NTM detection kit; PANAGENE, Korea) assay has been recently developed for the simultaneous detection of Mycobacterium tuberculosis complex (MTBC) and nontuberculous mycobacteria (NTM) in clinical specimens. The study was aimed at evaluation of the performance of PNA probe-based real-time PCR in respiratory specimens. METHODS: To evaluate potential cross-reactivity, the extracted DNA specimens from Mycobacterium species and non-mycobacterial species were tested using PNA probe-based real-time PCR assay. A total of 531 respiratory specimens (482 sputum specimens and 49 bronchoalveolar washing fluid specimens) were collected from 230 patients in July and August, 2011. All specimens were analyzed for the detection of mycobacteria by direct smear examination, mycobacterial culture, and PNA probe-based real-time PCR assay. RESULTS: In cross-reactivity tests, no false-positive or false-negative results were evident. When the culture method was used as the gold standard test for comparison, PNA probe-based real-time PCR assay for detection of MTBC had a sensitivity and specificity of 96.7% (58/60) and 99.6% (469/471), respectively. Assuming the combination of culture and clinical diagnosis as the standard, the sensitivity and specificity of the new real-time PCR assay for detection of MTBC were 90.6% (58/64) and 99.6% (465/467), respectively. The new real-time PCR for the detection of NTM had a sensitivity and specificity of 69.0% (29/42) and 100% (489/489), respectively. CONCLUSIONS: The new real-time PCR assay may be useful for the detection of MTBC in respiratory specimens and for discrimination of NTM from MTBC.