Urinary Orosomucoid (α-1-Acid Glycoprotein) As A Single Potential Biomarker Through PET Assay for Diagnosis of Early Onset Sepsis in Neonates: A Review

Neonatal sepsis is a condition in which bacteria are present in an infant’s sterile body fluids. It is considered one of the most common causes of infant death, with nearly one million deaths per birthday and approximately 2 million deaths in the first week of life. To aid in the early diagnosis of neonatal sepsis, a potential new biomarker for early neonatal sepsis called orosomucoid (ORM) or α1-glycoprotein (α1AGP) in urine is being evaluated because of its greater accuracy than current diagnostic tools. Combined with particle turbidity analysis (PET), neonatal sepsis can be diagnosed in an immediate, sensitive, specific and non-invasive manner. The early local increase in urinary ORM in sepsis suggests that it could be a new promising marker of sepsis and an important part of routine laboratory and clinical practice.

Off-label use of T2Candida® test in vitreous humor for diagnosing invasive candidiasis: a clinical report

Here, we present a case of off-label successful use of the T2 MR (T2Candida® test) for the diagnosis of invasive candidiasis ( Candida albicans endolphthalmitis). This case demonstrates that T2Candida could be performed in sterile body fluids to improve microbiological diagnosis of invasive candidiasis.

Applicability and performance of EUCAST’s rapid antimicrobial susceptibility testing (RAST) on primarily sterile body fluids in blood culture bottles in laboratory routine with total lab automation

Optimisation of microbiological diagnostics in primarily sterile body fluids is required. Our objective was to apply EUCAST’s RAST on primarily sterile body fluids in blood culture bottles with total lab automation (TLA) and to compare results to our reference method Vitek2 in order to report susceptibility results earlier. Positive blood culture bottles (BACTEC™ Aerobic/Anaerobic/PEDS) inoculated with primarily sterile body fluids were semi-automatically subcultured onto Columbia 5% SB agar, chocolate agar, MacConkey agar, Schaedler/KV agar and Mueller-Hinton agar. On latter, cefoxitin, ampicillin, vancomycin, piperacillin/tazobactam, meropenem and ciprofloxacin were added. After 6 h, subcultures and RAST were imaged and MALDI-TOF MS was performed. Zone sizes were digitally measured and interpreted following RAST breakpoints for blood cultures. MIC values were determined using Vitek2 panels. During a 1-year period, 197 Staphylococcus aureus, 91 Enterococcus spp., 38 Escherichia coli, 11 Klebsiella pneumoniae and 8 Pseudomonas aeruginosa were found. Categorical agreement between RAST and MIC was 96.5%. Comparison showed no very major errors, 2/7 (28.6%) and 1/7 (14.3%) of major errors for P. aeruginosa and meropenem and ciprofloxacin, 1/9 (11.1%) for K. pneumoniae and ciprofloxacin, 4/69 (7.0%) and 3/43 (5.8%) for Enterococcus spp. and vancomycin and ampicillin, respectively. Minor errors for P. aeruginosa and meropenem (1/8; 12.8%) and for E. coli and ciprofloxacin (2/29; 6.5%) were found. 30/550 RAST measurements were within area of technical uncertainty. RAST is applicable and performs well for primarily sterile body fluids in blood culture bottles, partially better than blood-based RAST. Official EUCAST evaluation is needed.

Antimicrobial Susceptibility of Bacteroides fragilis Group Isolates to Clindamycin, Tetracycline and Tigecycline, and Their Possession of tet and ermF Genes, which are Responsible for Resistance

Objective: We aimed to determine the resistance of Bacteroides fragilis group (BFG) bacteria to clindamycin, tetracycline and tigecycline and establish the distribution of related resistance genes. Method: In total 82 BFG strains, isolated from different clinical samples between January 2017 and December 2018, were identified by MALDI-TOF MS. Their antimicrobial sensitivities to were determined using agar dilution methodology (CLSI; M11-A7). The tetM, tetQ, tetX, tetX1, tet36, and ermF genes were investigated by PCR. Results: Eighty-two strains of BFG bacteria, isolated from intra-abdominal abscess (n=36), tissue biopsy (n=16), blood (n=14) and other sterile body fluids (n=12), were identified as Bacteroides fragilis (n=48), Bacteroides thetaiotaomicron (n=17), Bacteroides vulgatus (n=5), Bacteroides ovatus (n=4), Bacteroides caccae (n=1), Bacteroides uniformis (n=1) and Parabacteroides distasonis (n=6). The resistance rates to clindamycin, tetracycline and tigecycline were 54.9%, 84.1%, 4.9%, respectively. Non-B. fragilis isolates were more resistant than B. fragilis strains. In total, 57.3% of the isolates were ermF gene positive, while B. thetaiotaomicron had the highest rate (70.6%). The tet gene positivity ranged from 18.8% to 66.7% among species. The tetQ gene positivity was higher than other tet genes. The 92.7% of the isolates were resistant to at least one antibiotic, while 94% had at least one resistance gene. Conclusion: This study provided data on antimicrobial resistance of our BFG isolates to clindamycin, tetracycline and tigecycline and the related resistance genes. However, our information obtained could also be a starting point for further investigation of the antibiotic resistance mechanisms of Bacteroides species, as well as, resistance transfer among BFG isolates and other bacteria.

Superiority of a novel Mp1p antigen detection enzyme immunoassay compared to standard BACTEC blood culture in the diagnosis of talaromycosis

BACKGROUND Talaromycosis is an invasive mycosis endemic in Southeast Asia and causes substantial morbidity and mortality in individuals with advanced HIV disease. Current diagnosis relies on isolating Talaromyces marneffei in cultures, which takes up to 14 days and is detectable only during late-stage infection, leading to high mortality. METHODS In this retrospective case-control study, we assessed the accuracy of a novel Mp1p antigen-detecting enzyme immunoassay (EIA) in stored plasma samples of 372 patients who had culture-proven talaromycosis from blood or sterile body fluids(reference standard) and of 517 individuals without talaromycosis (338 healthy volunteers; 179 with other infections). All participants were recruited between 2011-2017 in Vietnam. RESULTS 66.1% and 75.4% of cases and controls were male; the median age was 33 and 37, respectively. All cases were HIV-infected; median CD4 count was 10 cells/mm3. At an optical density cut-off of 0.5, the specificity was 98.1% (95% CI: 96.3%-99.0%); the sensitivity was superior to blood culture, 86.3% (95% CI: 82.3%-89.5%) versus 72.8% (95% CI: 68.0%-77.2%), P<0.001, McNemar test. The time-to-diagnosis was 6 hours versus 6.6 ± 3.0 days for blood culture. Paired plasma and urine testing in the same patients (N=269) significantly increased sensitivity compared to testing plasma alone P<0.001, or testing urine alone P=0.02, McNemar tests. CONCLUSIONS The Mp1p EIA is highly specific and is superior in sensitivity and time-to-diagnosis compared to blood culture for the diagnosis of talaromycosis. Paired plasma and urine testing further increases sensitivity, introducing a new tool for rapid diagnosis, enabling early treatment and potentially reducing mortality.

Impact of Efflux Pump Inhibitor Carbonyl-Cyanide M-Chlorophenylhydrazone in Multidrug Resistant AcinetobacterSpecies Isolates from Sterile Body Fluids

Introduction: Antimicrobial resistance of Acinetobacter baumannii(A. baumannii) are rapidly emerging, becoming non-responsive to most of the commonly prescribed antibiotics and leaving us with few treatment options and galloping treatment costs. Aim: To study the effect of Efflux Pump Inhibitor (EPI) Carbonyl Cyanide 3-Chlorophenylhydrazone (CCCP) on Multidrug Resistance (MDR) A. baumannii isolates from different sterile body fluids. Materials and Methods: A total of 40 Acinetobacter species isolates from different sterile body fluids i.e., Cerebrospinal Fluid (CSF), ascitic fluid, pleural fluid, and peritoneal fluid were collected and identified by Matrix Assisted Laser Desorption/Ionisation-Time Of Flight (MALDI-TOF), Biomerieux, France. Minimum Inhibitory Concentration (MIC) of A. baumannii was determined by automated VITEK-2 Antimicrobial Susceptibility Testing (AST) system (Biomerieux, France). In addition, MIC of the isolates, grown on Mueller-Hinton Agar (MHA) plate with 15 μg/mL with EPI CCCP (Sigma Aldrich, US) was determined. For Tigecycline, MIC was determined by Broth Microdilution (BMD) method. Results: Out of 40 isolates, 34 (85%) were A. baumannii and 6 (15%) were Acinetobacter junii. Most of the Acinetobacter spps were MDR and only susceptible to few antibiotics. Most effective antibiotic was Tigecycline 25 (73.52%) followed by Co-trimoxazole 10 (29.41%). Similarly, Out of 40 isolates, 2 to 64 folds reductions in MIC was observed due to CCCP in 10 (25%) isolates for various antibiotics. Likewise, for Tigecycline, 2 to 4 folds reductions in MIC value (One strain changed from intermediate to sensitive) was observed by VITEK-2 AST which corroborated with reduction in MIC by BMD after addition of CCCP. Conclusion: MDR A. baumannii are spreading rapidly. There is the need to overcome the antimicrobial resistance by investigating resistance inhibiting substance that will help to restore antimicrobial susceptibility and bringing back the existing antibiotics in prescription.