Xpert Carba-R Assay on Flagged Blood Culture Samples: Clinical Utility in Intensive Care Unit Patients with Bacteremia Caused by Enterobacteriaceae.

Introduction and background: Rapid molecular diagnostics to predict carbapenem resistance well before the availability of routine drug sensitivity testing (DST) can serve as an antimicrobial stewardship tool in the context of high rates of Carbapenem-resistant Enterobacteriaceae (CRE). Materials and methods: A retrospective observational study of patients more than 18 years of age on whom Xpert Carba-R (FDA approved for rectal swab specimen) was done on gram-negative bacteria (GNB) flagged blood culture samples, in an Indian intensive care unit between January 2015 and November 2018. We analyzed the performance of Xpert Carba-R in comparison with routine DST. Results: A total of 164 GNBs were isolated from 160 patients. Klebsiella pneumoniae and Escherichia coli were the predominant isolates. Carba-R was positive in 35.36% of samples and 45.34% were carbapenem-resistant (CR) on routine DST. The distribution of the CR gene was: Oxacillinase (OXA) (50%), NDM (32.7%) followed by OXA and NDM co-expression (15.51%). The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, and negative predictive value of Carba-R were 90.74, 93.15, 13.25, 0.10, 83.58 and 96.31% for Enterobacteriaceae. The median time to obtain the Carba-R report was 30 hours 34 minutes vs 74 hours and 20 minutes for routine DST. Based on the Carba-R report, 9.72% of patients had escalation and 27.08% had de-escalation of antibiotics. Conclusion: Xpert Carba-R serves as a rapid diagnostic tool for predicting carbapenem resistance in intensive care unit patients with bacteremia caused by Enterobacteriaceae. How to cite this article: Rajendran S, Gopalakrishnan R, Tarigopula A, Kumar DS, Nambi PS, Sethuraman N, et al. Xpert Carba-R Assay on Flagged Blood Culture Samples: Clinical Utility in Intensive Care Unit Patients with Bacteremia Caused by Enterobacteriaceae. Indian J Crit Care Med 2023;27(9):655-662.

How Reliable Is Automated Urinalysis in Acute Kidney Injury?

OBJECTIVE: Examination of urine sediment is crucial in acute kidney injury (AKI). In such renal injury, tubular epithelial cells, epithelial cell casts, and dysmorphic red cells may provide clues to etiology. The aim of this study was to compare automated urinalysis findings with manual microscopic analysis in AKI. METHODS: Samples from patients diagnosed with AKI and control patients were included in the study. Red blood cells, white blood cells, renal tubular epithelial cells/small round cells, casts, and pathologic (path) cast counts obtained microscopically and by a UF1000i cytometer were compared by Spearman test. Logistic regression analysis was used to assess the ability to predict AKI from parameters obtained from the UF1000i. RESULTS: There was poor correlation between manual and automated analysis in AKI. None of the parameters could predict AKI using logistic regression analysis. However, the increment in the automated path cast count increased the odds of AKI 93 times. CONCLUSION: Automated urinalysis parameters are poor predictors of AKI, and there is no agreement with manual microscopy.

EGFR mutations and ROS1 and ALK rearrangements in a large series of non-small cell lung cancer in South India.

BACKGROUND: Driver mutations are seen in 80% of lung adenocarcinomas, and they influence prognosis and choice of therapy. AIM: Aim of this study was to analyse the frequency of epidermal growth factor receptor (EGFR) mutations, ALK and ROS1 rearrangements and their association with age and gender in non-small cell lung cancer reported from a tertiary care center in South India. METHODS: Tumors from patients with non-small cell carcinoma of lung were evaluated for EGFR mutations, ALK and ROS1 rearrangements and their association with age and gender were studied. RESULTS: Two thirds of non-small cell carcinomas had driver mutations or rearrangements. EGFR mutation was common and seen in 34.1%, whereas ALK rearrangement was seen in 11.1% and ROS1 rearrangement in 2% patients. Among EGFR mutations, most common were Exon 19 deletion and L858R seen in 21.3% and 11% of patients, respectively. Adenocarcinoma was the histologic diagnosis in 81% to 85% of patients with exon 19 deletion and L858R mutation, respectively. EGFR mutation frequency in patients less than 36 years was 13.6%, whereas in older patients, it varied from 34% to 36%. Exon 19 deletion was seen in 29.8% females and 17.2% of males. CONCLUSION: EGFR mutations are more common than ALK and ROS1 rearrangements. They are more common in females. Patients less than 36 years have reduced frequency of EGFR mutations. Exon 19 deletion and L858R are most common and are more prevalent in lung adenocarcinomas. Rare EGFR mutations are seen in patients aged more than 50 years.

Recurrent genetic abnormalities detected by FISH in adult B ALL and association with hematological parameters.

BACKGROUND: Recurrent genetic abnormalities influence prognosis in B lymphoblastic leukemia. BCR-ABL rearrangement is associated with higher leukocyte counts and older age at presentation. Among adults, BCR -ABL - is the commonest recurrent abnormality whereas, IgH rearrangements are rare. AIM: Aim of this study was to identify common recurrent genetic abnormalities in adult B ALL and study their association with hematological findings. METHODS: Bone marrow and peripheral blood from patients with B acute lymphoblastic leukemia were analyzed for complete blood counts, bone marrow morphology and cytogenetic abnormalities. The study group was divided into smaller groups based on cytogenetic abnormalities. Hematological parameters and presence of recurrent genetic abnormalities was compared across age groups and gender by non parametric tests. RESULTS: BCR-ABL positive group had a higher leukocyte count than BCR-ABL negative group. Among groups 1 to 5, group 1 with gains of chromosomes was associated with leucopenia and higher age at presentation. BCR-ABL is commonest recurrent abnormality followed by IgH rearrangements. CONCLUSION: Patients with gains of chromosomes alone have low total leukocyte counts at presentation.

Cytogenetic Abnormalities in Multiple Myeloma Patients at a Tertiary Healthcare Center in India

Objective: Multiple myeloma (MM) is a clinically and genetically heterogeneous plasma cell neoplasm. The prognosis of MM patients is dependent on several factors including the patient's age, the stage of disease and genetic alterations. This study aimed to determine the frequency of common chromosomal abnormalities and their significance in MM patients referred to a tertiary healthcare center in India. Methods: Fluorescence in situ hybridization on interphase nuclei from bone marrow cells using seven MM-specific probes for recurrent aberrations was performed in a total of 215 newly diagnosed patients. Results: Chromosomal abnormalities were detected in 161 (74.9%) MM patients in this study. The most frequent aberration was trisomy(ies) involving only gain of chromosomes in 48 (22.3%) cases. A translocation involving the IGH gene alone or accompanied by trisomy(ies) or by monosomy 13/13q deletion or by both was registered in 80 (37.2%) patients. Atypical patterns such as a deletion of the IGH variable segment (IGHv) on the derivative chromosome 14 or on the native (normal) chromosome 14, biallelic deletion of IGHv, deletion of the IGH constant segment on the rearranged chromosome14 and extra fusions were noticed in 21 (9.8%) patients with an IGH rearrangement. Monosomy 13/deletion 13q was identified singly or as part of a complex karyotype in 74 patients (34.4%). Clonal heterogeneity and additional abnormalities including TP53 deletion and monosomies of chromosomes 4, 9, 14 and 16 were recorded in 18.6% and 16.3% of patients respectively. Patients with abnormalities exhibited plasmacytosis, reduced hemoglobin value and high level of ß2-microglobulin. Conclusions: A lower median age and a low frequency of IGH translocations particularly t(11;14) and chromosome 13 abnormalities suggest ethnic diversity. Further investigations on genetic alterations including IGH deletions will contribute to improved insights into the biology of myeloma disease, risk stratification and patient management.

Can Platelet Distribution Width Be Used to Predict the Possibility of Chronic Myeloproliferative Neoplasms?

Platelet distribution width (PDW) and mean platelet volume are markers of platelet activation and have prognostic value in coronary heart diseases, as well as in cancers of solid organs. In this study, we evaluated the possibility of using PDW to predict chronic myeloproliferative neoplasms by comparing platelet indices obtained by automated analyzers in chronic myeloproliferative neoplasms with those in control specimens. We found that PDW greater than 66.4% has specificity of 99% and likelihood ratio of 19.5 for predicting chronic myeloproliferative neoplasms. Also, the area under curve (AUC) for platelet distribution width is 0.68.

Fatal aspergillosis of the renal vasculature in a combined liver-kidney transplant recipient.

Invasive aspergillosis remains a problem in solid organs and haematopoietic stem cell transplants. We report a case of 12-year-old female with primary hyperoxaluria with regular haemodialysis for the end-stage renal disease. She underwent a combined liver and renal transplantation which got infected by aspergillosis. In this case study, it is speculated that the most likely source of Aspergillus was contaminated preservative solution (perfusate), resulting in infection within the donor kidney and subsequent systemic infection in the recipient. This case study calls for critical analysis and needs for the routine culture of the preservative solution before transplantation, to detect any fungal contamination and manage it prophylactically.

Utility of multiplex polymerase chain reaction (PCR) in diarrhea-An Indian perspective.

BACKGROUND: Infective diarrhea causes morbidity worldwide. Polymerase chain reaction (PCR)-based pathogen diagnostics of diarrheal stool specimens are shown to be highly sensitive and rapid as opposed to conventional diagnostics. METHODS: We analyzed the performance of FilmArray gastrointestinal (GI) panel, one such multiplex PCR test, on stool specimens in patients presenting with diarrhea to our hospital from March 2016 to September 2017 and compared the results with conventional diagnostic tests. RESULTS: A total of 106 patients were included. The panel detected at least one target in 54 out of 106 patients (50.9%) with results available on the same day. Multiple targets were detected in 26 out of 54 patients who tested positive (48.1%). Bacteria as an isolated etiology for diarrhea was present in 34 patients (62.9%), viruses (16.7%, nine patients), parasites (7.4%, four patients), and multiple pathogens in seven patients (12.9%). Enteroaggregative Escherichia coli (EAEC) was the commonest pathogen detected (in 23, 24% patients). Conventional diagnostic investigations, undertaken in 68/106 (64.1%) patients were positive in 12 (17.65%) as compared to 54/106 (50.9%) (p < 0.0001). Conventional  investigations detected a pathogen not included in the study panel in 11 of 52 patients (21.1%). CONCLUSION: FilmArray multiplex PCR panel detects a wide array of GI pathogens including viruses and co-infections at a shorter time with more sensitivity compared to conventional diagnostics. Henceforth, it may facilitate treatment decisions, isolation policy, and antimicrobial stewardship in patients with diarrhea requiring hospitalization.

Cytogenetic Profiling of Myelomas, Association With Complete Blood Count: Study of 180 Patients.

OBJECTIVES: To analyze the most common primary and secondary cytogenetic events in myelomas using a probe panel designed in our laboratory, and to associate those events with hematological and biochemical findings. METHODS: Blood specimens from patients diagnosed with myeloma were processed to determine complete blood count and levels of albumin, creatinine, and beta-2 microglobulin. We evaluated bone-marrow specimens for plasma-cell percentage by light microscopy and for cytogenetic abnormalities by fluorescence in situ hybridization (FISH). The Mann-Whitney U test was used to compare hematological and biochemical parameters. RESULTS: We observed immunoglobulin heavy chain (IgH) gene translocations in 43.3% and t(4;14) in 21% of specimens; t(11;14) was observed in 7.7% of specimens. Gain of chromosomes was observed in 67.2% and loss observed in 16.6% of specimens. CONCLUSIONS: Gains of chromosomes were observed in two-thirds of patients with myeloma. The most common IgH translocation was t(4;14); del13/monosomy13 was the most common secondary cytogenetic abnormality. Partial or complete tetrasomies were associated with higher beta-2 microglobulin levels.

Outcome of allo simple limbal epithelial transplantation (alloSLET) in the early stage of ocular chemical injury.

PURPOSE: To analyse the outcome of allo simple limbal epithelial transplantation (alloSLET) in eyes in the early stage following ocular chemical injury to achieve rapid epithelialisation. PATIENTS AND METHODS: Records of 18 eyes of 17 patients who underwent alloSLET between April 2013 and Jan 2016 were analysed retrospectively. Patients with grade 4 or worse Dua's classification for chemical injury, who presented within a month of the injury or with a non-healing epithelial defect since the injury despite earlier medical or surgical interventions, were included in the study. The time to epithelialisation was the primary outcome measure and the best corrected visual acuity, the clinically assessed epithelial phenotype and symblepharon formation were the secondary outcome measures. RESULTS: The mean time to epithelialisation was noted to be 22.5±9.14 days. A best corrected visual acuity of better than 20/120 was achieved in 13 eyes in a mean duration of 33.06+10.73 days following alloSLET. Corneal phenotype with complete epithelialisation was achieved in the immediate postoperative period in 17 of the 18 eyes (94.11%). Seven eyes had a gradual failure of the allograft and 5 eyes underwent subsequent limbal autograft. Symblepharon formation involving one to two quadrants was noted in 3 eyes (16.7%). CONCLUSIONS: AlloSLET seems to be a useful technique to achieve rapid epithelialisation in severe chemical injuries thereby preventing the adverse effects of delayed epithelial healing. Visual rehabilitative procedures in the chronic phase of chemical injury, in most instances following an alloSLET in the acute stage, did not require keratoplasty, either lamellar or penetrating.