A retrospective analysis on seroprevalence of acute viral hepatitis observed among dengue patients attending a tertiary care centre in central India.

PURPOSE: The present study was conducted retrospectively to assess the frequency of acute viral hepatitis among the clinically suspected dengue cases presented at our tertiary care centre during 2021. METHODS: To determine the presence of acute viral hepatitis; Hepatitis A virus (HAV) and Hepatitis E virus (HEV) infections, 104 specimens were selected from the dengue-suspected clinical specimens received during 2021 on the basis of acute viral hepatitis symptoms. Following this, serological diagnosis was performed on those samples using anti-HAV IgM and anti-HEV IgM ELISA kits. RESULTS: Based on sero-positivity for IgM antibodies, 3 (5.3%) dengue virus (DENV) seropositive samples were positive for both HAV and HEV, while among DENV seronegative cases, 11 (22.91%) samples were positive for HEV and 1 (2.08%) sample was positive for HAV, pointing towards misdiagnosis due to overlapping symptoms. Additionally, co-infection of HAV & HEV in 1 sample was also observed in this study. CONCLUSIONS: This study revealed the presence of acute hepatitis infections among the dengue cases during monsoon and post-monsoon season. Overlapping of the clinical manifestations of these diseases can create misdiagnosis incidences raising risk for underreporting of the true cases of acute viral hepatitis infection. Dengue-suspected patients with selected symptoms during the monsoon and post-monsoon season should additionally be screened for acute hepatitis infections, as suggested in this study.

Performance Evaluation of Different RT-PCR Kits for the Direct Detection of SARS-CoV-2 in Preheated Specimens.

Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has created high demand for molecular kits and consumables for mass screening of suspected individuals. Direct real-time polymerase chain reaction (RT-PCR) assay without nucleic acid extraction has several advantages in saving testing time and cost and helps in the rapid reporting of SARS-CoV-2. The present study evaluated the analytical performance of four SARS-CoV-2 RT-PCR for direct RT-PCR testing using preheated specimens. Methods A total of 100 clinical specimens were selected and divided into three different groups: (1) group I: 20 SARS-CoV-2 positive specimens with high viral load, viz., low Ct values (< 30 Ct), (2) group II: 50 SARS-CoV-2 positive specimens with low viral load, viz., high Ct values (> 30 Ct), and (3) group III: 30 SARS-CoV-2 negative specimens. Specimens were heat-inactivated at 70°C for 10 minutes and cooled down at 4°C and were evaluated for standard and direct RT-PCR method by using ViralDtect-II Multiplex Real-Time PCR kit, TaqPath COVID-19 Combo kit, COVIDsure Pro Multiplex RT-PCR kit, and Hi-PCR Coronavirus (COVID-19) Multiplex Probe PCR kit. Results Results showed that except ViralDtect-II kit, the other three TaqPath COVID-19 Combo kit, COVIDsure Pro kit, and Hi-PCR Coronavirus (COVID-19) RT-PCR kit were able to amplify all the SARS-CoV-2 genes in the direct RT-PCR method using preheated specimens. In group I specimens, 100% sensitivity was observed in all three RT-PCR kits. In group II specimens, COVIDsure Pro kit was found to be superior among other kits. Conclusion Direct RT-PCR method during pandemic situation is valuable and cost effective for the detection of SARS-CoV-2. All three TaqPath COVID-19 Combo kit, COVIDsure Pro kit, and Hi-PCR Coronavirus (COVID-19) RT-PCR kit can be used for direct RT-PCR method and COVIDsure Pro kit performance was found to be superior among all.

Emergence of Multidrug-Resistant Non-Fermenting Gram-Negative Bacilli in a Tertiary Care Teaching Hospital of Central India: Is Colistin Resistance Still a Distant Threat?

Purpose Multidrug-resistant (MDR) organisms are being increasingly reported from India. This study aimed to determine the antibiotic susceptibility pattern of non-fermenting Gram-negative bacilli (NF-GNB) isolated from all the clinical samples to estimate the prevalence of MDR MDR NF-GNB and to screen for colistin-resistance genes among all colistin-resistant strains. Materials and methods This prospective study conducted from January 2021 to July 2022 at a tertiary care teaching hospital in central India identified MDR NF-GNB from clinical samples using standard procedures and antimicrobial susceptibility testing conducted as per Clinical Laboratory Standards Institute (CLSI) guidelines. Colistin-resistant strains identified by broth microdilution were further subjected to detection of plasmid-mediated colistin-resistant genes (mcr-1, mcr-2, mcr-3) by polymerase chain reaction (PCR). Results A total 2,106 NF-GNB were isolated from 21,019 culture positive clinical samples, of which 743 (35%) were MDR. Majority of MDR NF-GNB isolated were from pus (45.50%) followed by blood (20.50%). Out of 743 non-duplicate MDR non-fermenters,the most common were Pseudomonas aeruginosa (51.7%), Acinetobacter baumannii (23.4%),and others (24.9%).Around5.2% Pseudomonas aeruginosa and 2.3% Acinetobacter baumannii were resistant to colistin, and 88.2% were resistant to ceftazidime. Burkholderia cepacia complexwas 100% susceptible to minocycline and least susceptible to ceftazidime (28.6%). Out of 11, 10 (90.9%) Stenotrophomonas maltophilia were susceptible to colistin and least susceptible to ceftazidime and minocycline (27.3%). All 33 colistin-resistant strains (minimal inhibitory concentration ≥ 4 µg/mL) were found to be negative for mcr-1, mcr-2, and mcr-3 genes. Conclusion Our study showed a significantly wide variety of NF-GNB, ranging from Pseudomonas aeruginosa (51.7%), Acinetobacter baumannii (23.4%),to Acinetobacter haemolyticus (4.6%), Pseudomonas putida (0.9%), Elizabethkingia meningoseptica (0.7%), Pseudomonas luteola (0.5%), and Ralstonia pickettii (0.4%), which have not been commonly reported in literature. Of all the non-fermenters isolated in the present study, 35.28% were MDR, raising the concern for rationalizing antibiotic use and improving infection control measures to avert or slow the emergence of antibiotic resistance.

Stability of SARS-CoV-2 RNA in Viral Lysis Buffer Stored at Different Temperatures.

Objectives The present COVID-19 pandemic resulted in an increased need for molecular diagnostic testing. Delay in the specimen processing and suboptimal storage of suspected samples in laboratories leads to degradation of SARS-CoV-2 viral RNA. Viral lysis buffers from RNA extraction kits have the potential to stabilize RNA. Hence, this study aimed to investigate the stability of SARS-CoV-2 RNA in viral lysis buffer at different temperatures and time periods. Materials and Methods Aliquots of samples with known SARS-CoV-2 RNA were processed in viral lysis buffers simultaneously, stored separately at 2 to 8°C and 22 to 28°C for 24 hours, 48 hours and 72 hours. SARS-CoV-2 viral RNA was extracted from each aliquot and analyzed using multiplex real-time PCR. Results SARS-CoV-2 RNA in samples placed in viral lysis buffer was stable for 48 hours at both 2 to 8°C and 22 to 28°C temperatures. Slight decline in the viral RNA quantity was found on aliquots tested after 48 hours of both the temperatures. Conclusions Viral lysis buffer maintains the integrity of SARS-CoV-2 RNA for up to 48 hours even at room temperature and supports delayed diagnosis with an overwhelming sample load in testing laboratories.

Analysis of Antibiotic Resistance Genes and its Associated SCCmec Types among Nasal Carriage of Methicillin Resistant Coagulase Negative Staphylococci from Community Settings, Chennai, Southern India.

OBJECTIVE: The study was designed to find the distribution of SCCmec types and the various antibiotic resistance genes amongst MR-CoNS isolates from asymptomatic individuals. MATERIALS AND METHODS: A total of 145 nasal swabs were collected from asymptomatic healthy individuals from community settings. Identification and speciation of CoNS were done by standard biochemical methods. Screening of methicillin resistance (mecA gene) and detection of various antibiotic resistant genes were done using multiplex PCR method. SCCmec types (I - V) were determined using multiplex PCR. RESULTS: 50 (44.6%) isolates were found to be methicillin resistant both by cefoxitin method and multiplex PCR. S. epidermidis (40%) was the predominant species followed by S. haemolyticus (28%), S. hominis (20%) and S. warneri (12%). Highest resistance was shown for cotrimoxazole (26%), followed by ciprofloxacin (24%), tetracycline (20%), erythromycin (18%), fusidic acid (10%) and mupirocin (6%). Among SCCmec types, 44 isolates showed single type, including type I (30%), type IV (24%), type II (18%), type V (14%) and type III (2%). 6 isolates showed two types, III+IV (n= 2), II+V (n=2), IV+V (n=1) and type I+V (n=1). CONCLUSION: In conclusion, to the best of our knowledge, this is the first study in India to study the distribution of antibiotic resistant genes and SCCmec types among MR-CoNS from community settings. This study highlights high prevalence of MR-CoNS in community and its role in harbouring genetically diverse SCCmec elements as antibiotic resistance determinant.