Lodderomyces elongisporus fungemia in a patient with previous cardiac surgery: Case report and review of literature.

Lodderomyces elongisporus is a rare cause of invasive fungal infections. Most phenotypic tests that are routinely used for identification of yeasts fail to identify this organism. However, chromogenic media for yeasts, MALDI-TOF MS and DNA sequencing can be used for correct identification. We report a case of fungemia complicated by infective endocarditis and intracerebral bleeding in a pediatric patient with previous cardiac surgery.

Invasive pulmonary aspergillosis infection in severely ill COPD patients in pulmonary ward and ICU.

PURPOSE: This study was planned to determine the trends and susceptibility pattern of invasive pulmonary aspergillosis (IPA) in severely ill chronic obstructive pulmonary disease (COPD) patients admitted in pulmonary ward and ICU of our tertiary care centre. METHODS: Fifty COPD patients suspected of IPA from pulmonary ward and ICU from April 2017 to September 2018 were investigated. Samples were processed by standard methods, culture positive isolates were confirmed by MALDI-TOF MS and antifungal susceptibility testing was performed by microbroth dilution method. RESULTS: Twenty-two critically ill COPD patients were microbiologically positive for IA infection, of which 13 were classified as putative invasive aspergillosis. The most common comorbid illness associated was diabetes. A. flavus and A. fumigatus were the commonest species isolated. The minimum inhibitory concentration of the antifungals was low. Morbidity due to IPA in COPD patients was very high. CONCLUSIONS: Prevalence of IPA in the pulmonary ward and ICU was found to be 9.6%. MALDI-TOF seems to be a promising tool for aiding rapid identification especially for slow growing and non-sporulating fungi. Heightened awareness and suspicion for pulmonary mould infections along with early diagnosis can substantially alter the patient prognosis.

Gargle lavage & saliva: Feasible & cheaper alternatives to nasal & throat swabs for diagnosis of COVID-19.

Background & objectives: In the present scenario, the most common sample for diagnosis of COVID-19 by reverse transcription polymerase chain reaction (RT-PCR) is nasal and throat swab (NTS). Other sampling options such as gargle lavage have found limited application in clinical use mostly because of unavailability of an appropriate gargling liquid. This study was conducted to assess the stability of SARS-CoV-2 RNA in normal saline at 4°C that can serve as a gargling liquid as well as a transport medium. The study also looked at the agreement between NTS and gargle lavage/saliva for the detection of SARS-CoV-2. Methods: In 29 consecutive real-time RT-PCR (rRT-PCR) positive COVID-19 patients, paired NTS, gargle and saliva samples were taken. Samples were processed by rRT-PCR for the detection of SARS-CoV-2 RNA. To assess the SARS-CoV-2 RNA stability in normal saline, gargle lavage specimens were divided into two aliquots; one subset of the specimen was run within 4-6 h along with the routine samples (NTS and saliva) and the other subset was stored at 4°C and processed after 24-30 h. Agreement between cycle threshold (Ct) values from both the runs was compared using Bland-Altman (BA) analysis. Results: The positivity rates of rRT-PCR in NTS, saliva and gargle lavage samples were 82.7 (24/29), 79.3 (23/29) and 86.2 per cent (25/29), respectively. BA plot showed a good agreement between the Ct values of fresh and stored gargle samples, stipulating that there were no significant differences in the approximate viral load levels between the fresh and stored gargle lavage samples (bias: E gene -0.64, N gene -0.51, ORF gene -0.19). Interpretation & conclusions: Our study results show stability of SARS-CoV-2 RNA in the gargle samples collected using normal saline up to 24-30 h. Gargle lavage and saliva specimen collection are cost-effective and acceptable methods of sampling for the detection of SARS-CoV-2 RNA by rRT-PCR. These simplified, inexpensive and acceptable methods of specimen collection would reduce the cost and workload on healthcare workers for sample collection.

Development of a Sensitive and Specific Novel qPCR Assay for Simultaneous Detection and Differentiation of Mucormycosis and Aspergillosis by Melting Curve Analysis.

Molecular diagnostic assays can expedite the diagnosis of fungal infections, and subsequently help in early interventions and appropriate management of patients. The aim of this study was to develop a single set of primers for a real-time quantitative polymerase chain reaction (qPCR) assay to detect and identify commonly reported, clinically relevant molds i.e., Aspergillus spp, Mucorales and Fusarium spp., up to genus level by melting curve analysis. This assay was evaluated in whole blood from patients with suspected invasive aspergillosis (IA), and in tissue biopsy, bronchoalveolar lavage (BAL) fluid and other site-specific samples from patients with suspected invasive mucormycosis (IM). The limit of detection (LoD) was determined as 10 copies/µl for all three molds. The mean coefficient of variation (CV) across all sets of intra- and inter-assay data was 0.63% (ranging from 0.42 to 1.56%), showing high reproducibility of the assay. Sensitivity and specificity of the assay were 93.3 and 97.1% respectively for diagnosis of IA, and 99.29 and 83.84% respectively for diagnosis of IM. Fusarium was not detected in any of the clinical samples included and the few laboratory confirmed cases of fusariosis did not meet the inclusion criteria of the study. Hence no ROC curve or cutoff value could be generated for the same. This newly developed qPCR assay therefore appears to be a promising tool in detection of IA and IM.

SARS-CoV-2 RT-PCR profile in 298 Indian COVID-19 patients: a retrospective observational study.

BACKGROUND: despite being in the 5th month of pandemic, knowledge with respect to viral dynamics, infectivity and RT-PCR positivity continues to evolve. AIM: to analyse the SARS CoV-2 nucleic acid RT-PCR profiles in COVID-19 patients. DESIGN: it was a retrospective, observational study conducted at COVID facilities under AIIMS, New Delhi. METHODS: patients admitted with laboratory confirmed COVID-19 were eligible for enrolment. Patients with incomplete details, or only single PCR tests were excluded. Data regarding demographic details, comorbidities, treatment received and results of SARS-CoV-2 RT-PCR performed on nasopharyngeal and oropharyngeal swabs, collected at different time points, was retrieved from the hospital records. RESULTS: a total of 298 patients were included, majority were males (75·8%) with mean age of 39·07 years (0·6-88 years). The mean duration from symptom onset to first positive RT-PCR was 4·7 days (SD 3·67), while that of symptom onset to last positive test was 17·83 days (SD 6·22). Proportions of positive RT-PCR tests were 100%, 49%, 24%, 8·7% and 20·6% in the 1st, 2nd, 3rd, 4th and >4 weeks of illness. A total of 12 symptomatic patients had prolonged positive test results even after 3 weeks of symptom onset. Age > = 60 years was associated with prolonged RT-PCR positivity (statistically significant). CONCLUSION: this study showed that the average period of PCR positivity is more than 2 weeks in COVID-19 patients; elderly patients have prolonged duration of RT-PCR positivity and requires further follow up.

Mycobacterium abscessus Periprosthetic joint infection following bilateral Total Knee arthroplasty.

Periprosthetic joint infection (PJI) can be protracted, incapacitating, needing multiple interventions and could even lead to mortality. Early post-operative PJI has been ascribed to peri-operative introduction of highly virulent bacteria, while delayed post-operative to low-virulence bacteria. Non-tuberculous mycobacteria (NTM) do not figure in the usual list of etiological agents. We report a case of difficult diagnosis of bilateral PJI caused by Mycobacterium abscessus, following bilateral total knee arthroplasty in an elderly male, but treated successfully despite prolonged infection. M. abscessus complex comprises a group of rapidly growing, multidrug-resistant NTM, capable of forming biofilms on prostheses, responsible for wide spectrum of hospital acquired infections. M. abscessus as a cause of PJI is not reported widely. There are a few cases described in literature worldwide. There are no policy guidelines available for treating such cases. High clinical suspicion, with a concerted effort to grow and identify the causal pathogen is important. Standard anti-tubercular therapy is not recommended for treatment due to inherent resistance. Complete excision of infected tissues and removal of prosthesis along with prolonged combination antimicrobial regimen is the treatment of choice.

Mycobacterium fortuitum as a cause of acute CNS infection in an immune-competent girl undergoing repeated VP shunt surgeries.

We present the case of a 14-year-old immune-competent girl with ventriculoperitoneal shunt who was repeatedly hospitalised with meningeal signs despite repeated shunt revision surgeries. Eventually Mycobacterium fortuitum was isolated and the patient improved after specific treatment. M. fortuitum is a rapidly growing, non-tuberculous mycobacterium (NTM). NTMs are associated with postsurgical, post-trauma and device-related infections. Most of the present-day surgical equipment, catheters, prostheses and indwelling devices comprised silicone, stainless steel, polyvinyl chloride and polycarbonate, on which NTMs have the tendency to form biofilms. Central nervous system infection caused by NTM carries a high mortality rate (ranging from 35% to 70%), especially in immune-compromised patients. Indwelling device removal along with prolonged treatment with a combination regimen is recommended in such cases.