PURPOSE: Accurate identification of pathogens is critical for clinical microbiology laboratory. This helps in infection prevention and control and antimicrobial stewardship. The study objective was to develop and evaluate the importance of 16S ribosomal RNA sequencing for the identification of fastidious bacteria which are difficult to identify by conventional biochemical methods. This study also assessed the impact of this technology on rationalisation of antimicrobial therapy. MATERIALS AND METHODS: Clinical isolates or samples were chosen for 16S rRNA polymerase chain reaction (PCR) and Sanger sequencing based on the inability to detect or identify pathogens by conventional culture-based methods. This study was done on 148 patients for whom 16S rRNA PCR-Sanger sequencing was performed prospectively on clinical isolates/specimens from August 2015 to June 2017 in a tertiary cancer care hospital in Eastern India. Sanger sequencing was done using Applied Biosystems 3500 Genetic Analyzer. DNA sequence-based identification was done using NCBI BLAST. RESULTS: Forty-five different types (genera) of microorganisms were identified by this 16S rRNA PCR-sequencing method. Amongst the obligate aerobic bacteria identified by the 16S PCR sequencing method (n = 73) 55% were Gram positive and 45% Gram negative. Very few obligate anaerobes, capnophilic and microaerophilic bacteria were identified (n = 2, 1 and 6, respectively). Amongst facultatively anaerobic bacteria (n = 66), 64% were Gram negative and 35% Gram positive. The effect of 16S-based identification on antimicrobial stewardship has been documented. CONCLUSION: The study shows the potential importance of 16S rRNA-based bacterial identification as an important tool in clinical microbiology.

BACKGROUND: Since the prostate is in the area that is susceptible to infectious agents in Urine, it is rational to hypothesise that infectious agent that causes inflammation may be important in causing Prostate Cancer (PC). To date, there is no single microorganism that is recognised to aid in PC aetiology; yet, various species of bacteria are doubted or known to induce prostatitis, symptomatic bacterial prostatitis-related microbes and pro-inflammatory bacterium. MATERIALS AND METHODS: The left-over Urine samples were collected from patients diagnosed with carcinoma of the prostate attending the Urologic Clinic with a prior consent form after a histopathological diagnosis was performed. The Urine samples were streaked over petri plates (MacConkey and blood agar). Antibiotic susceptibility was conducted using the Kirby-Bauer disc diffusion method, following DNA extraction, polymerase chain reaction and 16s rRNA sequencing of the resistant isolates. IBM SPSS version 20.0 was used for the mean and standard error calculations with percentages. RESULTS: Urine samples showed the presence of polymicrobial isolates (85%), with Escherichia coli being the predominant organism (31.00%). Amongst 48 isolates, 23 were found resistant to antibiotic treatment. The sequencing analysis revealed that E. coli (7) and Klebsiella pneumonia (7) were predominant, followed by Enterococcus faecalis (5), Proteus vulgaris (2), Staphylococcus aureus (1) and Pseudomonas aeruginosa (1). CONCLUSIONS: The Urine analysis revealed polymicrobial isolates, indicating the presence of high microbial flora in men diagnosed with PC, indicating the prevalence of uropathogens and pro-inflammatory bacteria in the urinary tract of PC patients.

INTRODUCTION: Antibiotic resistance is a growing global health threat, especially when it comes to children. According to the WHO data, infections caused by multidrug-resistant bacteria produce 7,00,000 deaths across all ages, of which around 2,00,000 are newborns. This increasing trend in drug-resistant infections in the paediatric age group has gone unrecognised. It is essential to determine the trend of antibiotic resistance patterns of pathogenic bacteria isolated from the clinical samples for prompt and proper treatment of patients. The transmissible resistance spread through horizontal gene transfer and is associated with significant morbidity and mortality in the paediatric cases. Our aim of the study is antimicrobial resistance pattern of β-lactamase-producing bacteria isolated from children in a tertiary care hospital. OBJECTIVES: The objective of the study was to analyse the antimicrobial resistance pattern of the β-lactamase-producing organisms isolated from children. MATERIALS AND METHODS: The present study was a retrospective, observational, record-based one conducted in a selected hospital in Mangalore and data were collected from January 2019 to December 2021. All the clinical parameters, laboratory information and treatment histories were obtained from the patients case records and discharge summaries. Data were entered in MS Excel and analysed. RESULTS: A total of 2965 samples were studied; 757 cases were positive for culture. Extended-spectrum β-lactamase (ESBL) producers numbered 87 (11%) and metallo-β-lactamases (MBL) producers were 37 (5%). In our study, ESBL and MBL production was relatively high in uropathogens. In our findings, we have evidence showing increasing rates of ESBL and MBL producers in Gram-negative organisms amongst paediatric patients. CONCLUSION: Most of the study isolates were non β-lactamase producers; however, few percentages of Acinetobacter, Pseudomonas and Klebsiella spp. show both ESBL and MBL. We did not find MBL in E coli, Enterobacter, Proteus, Citrobacter and Stenotrophomonas isolates. Beta-lactamase production should be looked at routinely in Gram-negative bacteria. We recommend phenotypic identification methods as routine practice in laboratories.

A 59-year-old woman who had been undergoing chemotherapy for B-cell acute lymphoblastic leukaemia suffered from an episode of neutropenic sepsis on day 12 post-chemotherapy. Bacillus cereus (B. cereus) was isolated from two sets of blood cultures collected consecutively. The patient was treated with meropenem, colistin and teicoplanin. Although on the ventilator for three days, she recovered and could be discharged in 12 days. She continued to be on chemotherapy as per schedule. Bacillus spp. are usually considered contaminants in blood cultures but have been known to cause sepsis in immunocompromised patients. This case report aimed to highlight the possible pathogenic role of B. cereus when isolated from blood. Clinical correlation and routine practice of a minimum of two sets of blood cultures from two different sites will help us determine the significance of uncommon pathogens. The source of infection in this case is likely to be the gut, secondary to the mucosal barrier injury due to the chemotherapy.

This case report emphasises the significance of early diagnosis and treatment of Listeria monocytogenes infections as it can cause significant morbidity and mortality. Due to the limited/delayed diagnostic sensitivity of blood and cerebrospinal fluid (CSF) cultures, the diagnosis of Listeria meningitis is challenging. Here, we report the case of a 48-year-old male patient with multiple myeloma presented with fever, vomiting and headache. Routine investigations revealed hypocalcemia, elevated C-reactive protein and hypokalaemia. Despite being treated with ceftriaxone empirically, the patient's condition did not improve. Molecular testing of the CSF by the FilmArray meningitis/encephalitis panel identified L. monocytogenes within two hours. Ampicillin and Gentamicin were administered immediately and symptoms of the patient improved significantly. This case highlights the use of molecular syndromic panel like the FilmArray, which is highly sensitive for the diagnosis of central nervous system infections and enables targeted therapy and improved outcomes.

Candidaemia due to Candida spp. is a major cause of morbidity and mortality in the healthcare settings. Candida tropicalis is a globally emerging opportunistic pathogen causing invasive infection in immunocompromised patients. It is the most common non-albicans candida causing infections in humans. Clinical manifestations may vary from simple oral thrush to widely disseminated systemic infections, especially in immunocompromised patients. We report a case of an eight-year-old child with B-cell acute lymphoblastic leukaemia who developed widely disseminated candidiasis caused by Candida tropicalis with oral, dermatologic, hepatic and renal manifestations post-chemotherapy. The patient was empirically started on tablet Fluconazole and Amphotericin B was added after antifungal susceptibility testing. Good clinical response was seen after she was treated with injection Caspofungin.

More than 80% of instances of primary psoas abscess are caused by Gram-positive microorganisms. Our case, a 52-year-old man was admitted with one month history of back pain and fever. Computed tomography revealed a hypodense collection in the psoas muscle. Blood cultures for Brucella spp were positive approximately after three days and became negative, as the specific therapy was started. Complete resolution of symptoms was achieved within six weeks. Early aetiological identification is crucial due to the high success rate with appropriate antibiotic regimens, even though the clinical presentation of psoas abscesses due to different pathogens is frequently identical and symptoms are non-specific. Although Brucellosis is endemic in India, Brucellar psoas abscess appears to be rare. In this report, we describe a patient who had atypical symptoms, and blood culture helped to solve the diagnostic conundrum.

The objective of the study was to do galactomannan testing in the serum and BAL samples and correlate it with clinical data. The galactomannan testing was done by an immunoenzymatic sandwich microplate assay in 49 patients. Clinical data could not be obtained for nine patients. In the remaining 40 patients, single serum or BAL galactomannan was positive in 14 patients (35%) who had invasive pulmonary aspergillosis (IPA), and galactomannan was negative in 26 patients (65%) who had non-invasive pulmonary aspergillosis (non-IPA). Galactomannan tests were positive in 4 serum samples and 10 BAL samples. The test was negative in 15 serum samples and 11 BAL samples. BAL was the better sample for galactomannan testing, with a p-value of 0.048. Among the 14 patients with IPA and galactomannan positivity, only 2 had growth of Aspergillus, whereas among the 26 non-IPA patients with galactomannan negativity, only one grew Aspergillus. There is statistical significance in fungal culture between the two groups (p-value 0.023). The number of patients undergoing hemodialysis in the IPA group was 3, and all were positive for the galactomannan assay. In the non-IPA group, one patient had undergone hemodialysis and was negative for the galactomannan test. There is statistical significance between the two groups (p-value 0.027). The galactomannan assay must be carefully interpreted along with the clinical radiological findings, type of treatment, and associated cross-reactions.