Positive Blood Samples Research Articles

Common Contaminants Of Blood Group Factors And Its Effect On Blood Groups In Blood Samples, Collected At Scene Of Crime

For detection of crime in Forensic medicine blood group studies& its application plays an important role. The study of blood grouping gives important information regarding identification and details of Crime Investigation. The most common trace evidence is blood which is obtained from the crime scene. Blood grouping & typing can point to a group of people that have the same blood type rather than pointing particular person. When blood is properly collected and preserved, blood as an evidence can establish a strong link between a criminal and the criminal act. We can trust the blood grouping of blood stains in detection of crime. Randomly A, B, AB, Rh positive blood samples were taken in duplicates and rusted pins, dust particles, detergent, staphylococci are added to it respectively. The common contaminants in blood stains are dust, rust, bacteria and detergent. So an effect of these on blood groups was studied in detail.

Molecular detection of Coxiella burnetii in blood and hard tick-infested Egyptian camels and the possibility of coinfections

Coxiella burnetii, a bacterium that causes Q fever. It can infect mammals and has a global geographical distribution, but data on its occurrence in Egyptian dromedaries and the associated ticks are limited. Therefore, this study aims to detect C. burnetii in the blood of infested camels and associated ticks collected from Egypt by using molecular techniques and to examine the possibility of coinfections with C. burnetii. A total of 133 blood samples and 1260 hard ticks infesting these camels were collected from Egyptian slaughterhouses. Nested PCR and sequencing were used based on the IS1111 gene for molecular detection of C. burnetii. The identification of tick species at the molecular level was performed using the COX1 gene. C. burnetii was detected in Hyalomma (H) dromedarii, H. anatolicum, H. marginatum, Amblyomma (Am) lipidium, and Am. cohaerens with an overall prevalence rate of 1.3% (16/1260), while in the camel blood samples, it was 15.8% (21/133). Out of C. burnetii-positive ticks, there were double infections by Borrelia species and C. burnetii in H. dromedarii and Am. lipidium and triple infections at one Am. cohaerens tick (C. burnetii, Borrelia spp., and Babesia microti). In addition, two positive camel blood samples were found to carry C. burnetii with Borrelia spp. Our research findings indicate the presence of Coxiella burnetii among camels and their associated ticks in Egypt and emphasize the potential of having coinfection. To prevent the transmission of this infection to other animal species or humans, appropriate control measures should be implemented.

Development and validation of a new and rapid molecular diagnostic tool based on RT-LAMP for Hepatitis C virus detection at point-of-care

PurposeGlobally, it is estimated that 1.0 million individuals are newly infected by Hepatitis C virus (HCV) every year, and nearly 50 million people live with a chronic infection, according to World Health Organization. To overcome underdiagnosis of HCV infection among hard-to-reach populations, it is essential to develop new rapid and easy-to-use molecular diagnostic systems. In this work, we have developed a pangenotypic diagnostic tool based on Loop-Mediated Isothermal Amplification (LAMP), coupled to a direct sample lysis procedure for molecular detection of HCV at point-of-care (POC). MethodsProcedure validation was performed using 129 different samples from HCV infected patients (116 serum samples, and 13 fresh blood samples), 27 individuals who tested negative for HCV but positive for HIV, and 11 healthy donors. Serum was collected, lysed for 10 min at room temperature, and assayed by RT-LAMP. To achieve this, a set of 9 LAMP-primers was used for the first time. Parallel RT-qPCR assays were conducted for HCV to both validate the procedure and quantify viral loads. ResultsHCV was detected by RT-LAMP in 109/116 HCV positive serum samples, and in 11/13 positive blood samples in less than 40 min. Compared to RT-qPCR results, our RT-LAMP procedure showed a sensitivity of 94 %, 100 % specificity, and a limit of detection of 3.26 log10 IU/mL (10–20 copies per reaction). ConclusionsWe have developed an accurate system, more affordable than the current available rapid tests for HCV. Since no prior RNA purification step from capillary blood is required, we strongly recommend our RT-LAMP system as a valuable and rapid tool for the molecular detection of HCV at POC.

Molecular characterisation of hepatitis A in the Western Cape province, South Africa in 2023

In 2023, passive laboratory-based surveillance showed an increase in hepatitis A virus (HAV) infection. We investigated hepatitis A incidence using the notifiable medical condition surveillance system (NMCSS) data and molecularly characterised positive blood samples from the Western Cape province for 2023. All HAV IgM seropositive cases from the NMCSS from 1 January to 31 October 2023 in South Africa were investigated. HAV RNA from blood samples that had tested positive for HAV IgM from Western Cape was amplified in the VP1/P2B junction and sequenced (3500Xl Genetic Analyser). Sequences were assembled, aligned (Sequencher) and analysed (Aliview 1.27 and MEGA11). Statistical analysis was performed using Excel and the CuSum2 Threshold to determine suspected outbreaks. In 2023, the incidence of HAV IgM was 6.28/100,000 in South Africa, with the highest incidence in Western Cape province (15.86/100,000). Children aged 5 to 14 years were affected the most in the Western Cape. The positive cases in the Western Cape were above the CuSum2 threshold from January to May 2023, with the highest incidence observed in the City of Cape Town Metropolitan (14.8/100,000). Genotyping was successfully performed on 92.7% (139/150) of serum samples, for which the IB sub-genotype was detected. Three primary mutations R63K, R71S and M104I were observed in more than 49% of the samples. Most of the samples sequenced belonged to patients residing in areas close to each other within the City of Cape Town Southern, Western, and Mitchells Plain sub-districts. The CuSum2 threshold method allowed the identification of suspected HAV outbreaks in the districts within the Western Cape in 2023 while genotyping identified clusters of sub-genotype IB. Genotyping could assist with determining the common source of infection during an outbreak, especially if coupled with epidemiological and geographical data. Further active surveillance can assist in investigating the HAV risk factors for targeted public health responses.

Insights into tick dynamics and anaplasmosis in ruminants: A microscopic and molecular perspective

AbstractTicks are blood‐sucking parasite and transmit pathogens to humans, domestic and wild animals. Ticks are seriously damage the production of sheep and goats, resulting in heavy economic loss to farmers. The present study investigated microscopic and molecular identification of ticks and tick‐borne anaplasmosis infesting sheep and goats in three agro‐ecological zones of Pakistan. The collected ticks were identified at species level based on their morphological features through microscopy by using taxonomic keys. Four Rhipicephalus (R.) species namely: R. guilhoni Rh. geigyi, R. annulatus, R. pulchellus, two Haemaphysalis (Hae) spp. Hae. sulcate, Hae. punctate, one Ixodes ricinus and Hyalomma anatolicum were identified. The morphological variations in their body sizes, scutal pattern, female genital opening, spiracular plate length and male adanal plate were recorded. Furthermore, we assessed the effectiveness of utilizing the 16S rRNA gene sequence to differentiate Anaplasma samples at the species level. Microscopically suspected positive blood samples for Anaplasma were subjected to DNA extraction. Anaplasma was observed in the blood of 14 animals. The sequence similarity between Anaplasma marginale and the reference genome were 99.25%, while below this value indicating a distinct species. Phylogenetic analysis revealed close similarity with A. marginale isolates from Iran, Vietnam and Pakistan This study contributes to the epidemiology of ticks and TBPs in the KP province and highlights the importance of proactive TBP surveillance in livestock.

Blood and sputum microbiota composition in Afghan immigrants and Iranian subjects with pulmonary tuberculosis.

TB infection is one of the most challengeable epidemiological issues. Complex interactions between microbiota and TB infection have been demonstrated. Alteration in microbial population during TB infection may act as a useful biomarker. The present study examined the microbiota patterns of blood and sputum samples collected from Afghan immigrants and Iranian patients with active TB. Sixty active pulmonary TB patients were enrolled in the study. Blood and sputum samples were collected. To detect phylum bacterial composition in the blood and sputum samples, bacterial 16S rRNA quantification by Real-Time qPCR was performed. A significant decrease in Bacteroidetes in Iranian sputum and blood samples of Afghan immigrants and Iranian TB active subjects were seen. While, sputum samples of Afghan immigrants showed no significant differences in Bacteroidetes abundance among TB active and control. Firmicutes were also presented no significant difference between sputum samples of the two races. Actinobacteria showed a significant increase in Iranian and Afghan sputum samples while this phylum showed no significant abundance in Iranian and Afghan TB positive blood samples. Proteobacteria also showed an increase in sputum and blood samples of the two races. An imbalance in Bacteroidetes and Firmicutes abundance may cause an alteration in the microbiota composition, resulting in dysregulated immune responses and resulting in the augmentation of opportunistic pathogens during TB infection, notably Proteobacteria and Actinobacteria. Evaluation of human microbiota under different conditions of TB infection can be critical to a deeper understanding of the disease control.

The Diagnostic Accuracy of Procalcitonin and Its Combination with Other Biomarkers for Candidemia in Critically Ill Patients.

Background: The aim of this study was to investigate the usefulness of serum procalcitonin (PCT), C-reactive protein (CRP), neutrophil to lymphocyte count ratio (NLR), and their combination, in distinguishing candidemia from bacteremia in intensive care unit (ICU) patients. Methods: This is a retrospective study in ICU patients with documented bloodstream infections (BSIs) and with both serum PCT and CRP measurements on the day of the positive blood sample. Illness severity was assessed by sequential organ failure assessment (SOFA) score on both admission and BSI day. Demographic, clinical, and laboratory data, including PCT and CRP levels and NLR on the day of the BSI, were recorded. Results: A total of 63 patients were included in the analysis, of whom 32 had bacteremia and 31 had candidemia. PCT, CRP, and NLR values were all significantly lower in candidemia compared with bacteremia (0.29 (0.14-0.69) vs. 1.73 (0.5-6.9) ng/mL, p < 0.001, 6.3 (2.4-11.8) vs. 19 (10.7-24.8) mg/dl, p < 0.001 and 6 (3.7-8.6) vs. 9.8 (5.3-16.3), p = 0.001, respectively). PCT was an independent risk factor for candidemia diagnosis (OR 0.153, 95%CI: 0.04-0.58, p = 0.006). A multivariable model consisting of the above three variables had better predictive ability (AUC-ROC = 0.88, p < 0.001), for candidemia diagnosis, as compared to that of PCT, CRP, and NLR, whose AUC-ROCs were all lower (0.81, p < 0.001, 0.78, p < 0.001, and 0.68, p = 0.015, respectively). Conclusions: A combination of routinely available laboratory tests, such as PCT, CRP, and NLR, could prove useful for the early identification of ICU patients with candidemia.

Serotype and genomic diversity of dengue virus during the 2023 outbreak in Pakistan reveals the circulation of genotype III of DENV-1 and cosmopolitan genotype of DENV-2.

Dengue, a mosquito-borne viral disease, poses a significant public health challenge in Pakistan, with a significant outbreak in 2023, prompting our investigation into the serotype and genomic diversity of the dengue virus (DENV). NS-1 positive blood samples from 153 patients were referred to the National Institute of Health, Pakistan, between Julyand October 2023. Among these, 98 (64.1%) tested positive using multiplex real-time PCR, with higher prevalence among males (65.8%) and individuals aged 31-40. Serotyping revealed DENV-1 as the predominant serotype (84.7%), followed by DENV-2 (15.3%). Whole-genome sequencing of 18 samples (DENV-1 = 17, DENV-2 = 01) showed that DENV-1 (genotype III) samples were closely related (>99%) to Pakistan outbreak samples (2022), and approx. > 98% with USA (2022), Singapore and China (2016), Bangladesh (2017), and Pakistan (2019). The DENV-2 sequence (cosmopolitan genotype; clade IVA) shared genetic similarity with Pakistan outbreak sequences (2022), approx. > 99% with China and Singapore (2018-2019) and showed divergence from Pakistan sequences (2008-2013). No coinfection with dengue serotypes or other viruses were observed. Comparisons with previous DENV-1 sequences highlighted genetic variations affecting viral replication efficiency (NS2B:K55R) and infectivity (E:M272T). These findings contribute to dengue epidemiology understanding and underscore the importance of ongoing genomic surveillance for future outbreak responses in Pakistan.

Molecular markers of artemisinin resistance during falciparum malaria elimination in Eastern Myanmar

BackgroundArtemisinin resistance in Plasmodium falciparum threatens global malaria elimination efforts. To contain and then eliminate artemisinin resistance in Eastern Myanmar a network of community-based malaria posts was instituted and targeted mass drug administration (MDA) with dihydroartemisinin-piperaquine (three rounds at monthly intervals) was conducted. The prevalence of artemisinin resistance during the elimination campaign (2013–2019) was characterized.MethodsThroughout the six-year campaign Plasmodium falciparum positive blood samples from symptomatic patients and from cross-sectional surveys were genotyped for mutations in kelch-13—a molecular marker of artemisinin resistance.ResultThe program resulted in near elimination of falciparum malaria. Of 5162 P. falciparum positive blood samples genotyped, 3281 (63.6%) had K13 mutations. The prevalence of K13 mutations was 73.9% in 2013 and 64.4% in 2019. Overall, there was a small but significant decline in the proportion of K13 mutants (p < 0.001). In the MDA villages there was no significant change in the K13 proportions before and after MDA. The distribution of different K13 mutations changed substantially; F446I and P441L mutations increased in both MDA and non-MDA villages, while most other K13 mutations decreased. The proportion of C580Y mutations fell from 9.2% (43/467) before MDA to 2.3% (19/813) after MDA (p < 0.001). Similar changes occurred in the 487 villages where MDA was not conducted.ConclusionThe malaria elimination program in Kayin state, eastern Myanmar, led to a substantial reduction in falciparum malaria. Despite the intense use of artemisinin-based combination therapies, both in treatment and MDA, this did not select for artemisinin resistance.

Characterization of mutations in hepatitis B virus DNA isolated from Japanese HBsAg-positive blood donors in 2021 and 2022

Missense mutations in certain small envelope proteins diminish the efficacy of antibodies. Consequently, tracking the incidence and types of vaccine-escape mutations (VEMs) was crucial both before and after the introduction of universal hepatitis B vaccination in Japan in 2016. In this study, we isolated hepatitis B virus (HBV) DNA from 58 of 169 hepatitis B surface antigen (HBsAg)-positive blood samples from Japanese blood donors and determined the nucleotide sequence encoding the small envelope protein. DNA from six (10%) of the samples had VEMs, but no missense mutations, such as G145R, were detected. Complete HBV genome sequences were obtained from 29 of the 58 samples; the viral genotype was A1 in one (3%), A2 in three (10%), B1 in nine (31%), B2 in five (17%), B4 in one (3%), and C2 in 10 (34%) samples. Tenofovir-resistance mutations were detected in two (7%) samples. In addition, several core promoter mutations, such as 1762A>T and 1764G>A, and a precore nonsense mutation, 1986G>A, which are risk factors for HBV-related chronic liver disease, were detected. These findings provide a baseline for future research and highlight the importance of ongoing monitoring of VEMs and drug resistance mutations in HBV DNA from HBsAg-positive blood donors without HBV antibodies.

Role of direct antiglobulin test in anemia under evaluation: Its prevalence, laboratory workup, and significance in further patient management-A study from eastern India.

The direct antiglobulin test (DAT) is an important diagnostic tool for immune hemolytic anemia (IHA). The present study was primarily aimed to identify the prevalence of DAT positivity in anemia patients along with its specificity . A three months follow up of the DAT positive patients were performed for the response during course of illness in terms of transfusion requirement, hemoglobin level, persistence of DAT. This cross sectional study was performed at a government medical college on symptomatic anemia patients. At initial evaluation, complete blood count (CBC), blood grouping and DAT were performed in the EDTA blood. DAT positive blood samples were analyzed for their immunoglobulin specificity, auto or alloantibody type. Acid elution and red cell phenotyping were performed wherever applicable.Their clinical presentation, hematological and biochemical parameters of hemolysis were evaluated. Statistical analysis was performed on the results on SPSS (Version 23.0;.USA) and Graph pad Prism version 9. P value <0.05 was considered significant. DAT was present in 64 out of 501 patients with male female ratio 1: 4. Warm AIHA (WAIHA) was 93.7% with secondary WAIHA 60%. IgG was associated in 86% DAT positive samples, Only C3d was 14%. All the 4 cold AIHA (6.3%) had a higher antibody titre and thermal amplitude. DAT strength was directly proportional to the degree of hemolysis. During 3 months follow up , persistence of DAT and blood transfusion requirement was more in secondary WAIHA . Hemoglobin increment was more in primary WAIHA (75%). DAT played a significant role in the diagnosis as well as evaluation of AIHA.

Molecular detection and identification of Plasmodium spp. isolated from captive-bred cynomolgus monkeys in Bogor, Indonesia

Background and Aim: Asian macaques are natural hosts of several Plasmodium species. Some monkey malaria parasites may infect humans and cause zoonotic infections. This study was conducted to estimate the prevalence of monkey malaria parasites in Bogor, Indonesia, based on molecular detection and identification, particularly in cynomolgus monkeys, which have a wide geographic distribution and share extensive habitats with humans. These data are needed to evaluate the status of simian malaria among macaques in Bogor and to study the potential risks to human health. These updated data will provide sufficient information for implementing malaria control strategies in the future and for developing a potential malaria vaccine using monkeys as an animal model. Materials and Methods: Blood samples of 274 cynomolgus monkeys (Macaca fascicularis) were collected and identified using microscopy. DNA was extracted from positive blood samples and analyzed using polymerase chain reaction (PCR) to amplify the small subunit ribosomal RNA (SSU rRNA) target gene using consensus primers for Plasmodium species. The PCR-positive samples were then nucleotide-sequenced using commercial sequencing services, analyzed using the BioEdit program, and aligned using Basic Local Alignment Search Tool from the National Center for Biotechnology Information. Phylogenetic trees were constructed using MEGA 11.0 and the neighbor-joining (NJ) method to determine the kinship of Plasmodium. Bootstrapping was performed using 500 replicates to assess the robustness of tree topologies. Results: Thirty-eight of the 274 microscopically positive samples for Plasmodium spp. were also positive using PCR, resulting in a 1640 bp amplicon. Further, analysis using nucleotide sequencing confirmed that these positive samples were Plasmodium inui with more than 99% nucleotide identity compared to GenBank sequences. Phylogenetic tree analysis of the SSU rRNA partial gene showed that all our isolates clustered and were closely related to a P. inui strain isolated from cynomolgus macaques in South China in 2011. Conclusion: P. inui is the predominant malaria parasite in cynomolgus monkeys from Bogor. Keywords: malaria, phylogenetic tree, Plasmodium inui, small subunit ribosomal RNA.

Molecular Epidemiology of Plasmodium falciparum Infections Using PCR-based Assays in Jos, Nigeria-cross-sectional Study

Background: Malaria remains a significant health threat globally, with Plasmodium falciparum being the predominant and lethal parasite in Africa. Nigeria is still faced with ongoing cases of asymptomatic malaria, hindering effective control measures.&#x0D; Aim: The aim was to generate epidemiological data that will provide good background and guide strategies for driving malaria control efforts, research, and resource allocation in the region.&#x0D; Place and Duration of Study: This study was conducted in Jos, Plateau State, Nigeria, where the samples were originally collected within about 16 months between October 2019 and January 2021.&#x0D; Methodology: A cross-sectional molecular epidemiological study was conducted using 136 microscopically screened 2 plus (++) and above positive malarial whole blood samples obtained in EDTA bottles from two hospitals in Jos, Plateau State, Nigeria. The DNA extraction was performed according to the manufacturer's instructions using Zymo Research extraction kits. Plasmodium genus and Plasmodium falciparum were detected in the samples using the PCR method and gel electrophoresis.&#x0D; Results: In the results, using PCR techniques, 47.8% (65/136) of the total malaria-positive samples collected were confirmed for the presence of the Plasmodium genus. Out of these 65 positive samples, 63 were found to be Plasmodium falciparum.&#x0D; Conclusion: This study demonstrates that Plasmodium falciparum remains the predominant malaria species in Jos, Plateau State, comprising approximately 96.9% (63/65) of the malarial cases. This indicates that only about 3% of malaria cases affecting the residents of Jos, Plateau State might be caused by the other four species of malaria parasites (Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, and Plasmodium knowlesi).

Gambaran Residu Paparan Formalin pada Sampel Plasma Darah Karyawan Instalasi Hemodialisis Rumah Sakit

Hemodialysis is one of the installations in the hospital that handles blood washing therapy, by removing excess waste or harmful compounds, through a semi-permeable membrane which is carried out to replace kidney function that is no longer functioning properly. Hemodialysis is given to patients with a diagnosis of acute renal failure, terminal renal failure and patients who have severe poisoning with certain drugs. The research design was carried out descriptively. The aim of this research was to determine whether there was formalin exposure to hemodialysis workers or nurses when reusing or cleaning hemodialysis equipment, using a qualitative test. In this research, the results were obtained from 15 blood plasma samples, namely 6 positive formalin blood samples, with Schiff, Tollens, KMnO4 and Fehling reagents. It is known from the table of analysis results of qualitative test of formalin exposure on hemodialysis workers in one of the East Java Hospitals. For further research, it is hoped that quantitative tests will be carried out in determining the level of formalin in the blood using the instrument.

PCR combined with lateral flow dipstick assay (PCR-LFD) for a rapid diagnosis of melioidosis.

Melioidosis is an infectious disease caused by Burkholderia pseudomallei. Septicemic melioidosis patients have a high mortality rate within 48 hours. To develop a polymerase chain reaction (PCR) combined with a lateral flow dipstick (LFD) assay for detection of B. pseudomallei in blood samples. The PCR with wcbG gene primers and a PCR-LFD test were developed. The specificity and sensitivity were determined using the B. pseudomallei and other bacterial DNAs. They were evaluated using 43 B. pseudomallei positive blood samples and another 43 blood samples positive for other microbial infections. The detection limit of the PCR-LFD test was 50 fg of bacterial gDNA or 1.0 CFU per 200 μl of blood. All B. pseudomallei were positive while B. thailandensis and selected gram-negative bacterial strains were negative. The PCR-LFD gave all positives with all 43 B. pseudomallei culture positive patient blood samples and all negative with 43 blood samples that were culture positive for K. pneumoniae, E. gallinarum, E. faecium, E. coli, S. aureus, A. baumannii, A. hydrophila, S. haemolyticus, S. pneumoniae, P. aeruginosa, E. cloacae, S. hominis, E. aerogenes, P. mirabilis, C. neoformans, C. albicans, A. caviae, E. faecalis and K. variicola. The developed PCR-LFD assay provided 100% sensitivity and 100% specificity compared to the conventional blood culture. The technique took only 1.5 hours that is easy and quick to perform compared to the 3-7 days of culture method. The new method of PCR with LFD could facilitate the detection to be a semi-point-of-care testing (POCT).

Non-Synonymous Mutations Associated with Plasmodium falciparum Artemisinin Resistant Gene (Pfkelch13) in Malaria Cases, Jos Nigeria

Aim: The aim of this study was to explore the Plasmodium falciparum Artemisinin Resistant Gene (Pfkelch13) in malaria cases in Jos and profile the nature of the mutations. The Plasmodium falciparum kelch13 gene is a potential molecular marker for tracking artemisinin-resistant malaria parasites. Study Design: The Study Design was Experimental Place and Duration of Study: The study was conducted within Jos, Nigeria between October 2019 and January 2021. Methodology: Thirty-six clinically screened 2 plus (++) and above positive malarial whole blood samples were collected from a Hospital in Jos, Plateau State, Nigeria in EDTA bottles after being granted ethical approval. The DNA extraction was done using Zymo Research extraction kits according to the manufacturer’s instructions. Detection of the Plasmodium genus, Plasmodium falciparum and Pfkelch13 gene in the samples was done using PCR technique and gel electrophoresis. PCR amplicons were sequenced and bioinformatics software was used to analyze the sequences for mutations. Results: Only 41.67% of the samples collected were confirmed positive for the Plasmodium genus. Out of these, 93.33% were positive for Plasmodium falciparum. The PfKelch13 (Pfk13) gene was detected in 78.57% of the Plasmodium falciparum. Non-synonymous mutations (S695C, C696M, C696H, H697S, H697V, F698I, 699L and 699S were observed at the amino acids level, but didn’t affect the structural conformation of the proteins. Conclusion: The classical method of detecting malaria is not reliable compared to the PCR technique. Non-synonymous mutations on the PfKelch13 gene have the potential to cause resistance to artemisinin drugs in the Jos human population.

Screening of hepatitis B and C viral infection, recognition of risk factors, and immunization of patients against hepatitis B virus: a module developed for effective hepatitis control.

The continually increasing incidence of hepatitis, a worldwide health issue, in Pakistan, has highlighted the need to investigate the epidemiology factors and implement preventive measures accordingly. The purpose of this study was to scrutinize the prevalent and significantly associated risk factors of hepatitis in students and employees, screening them for hepatitis B and C virus and vaccinating them against HBV to make IUB hepatitis free. A total of 12,912 participants including students (n = 10,948) and employees (n = 1964) were screened for HBV and HCV via immunochromatographic test. Hepatitis- positive participants' blood samples were further tested and viral load was estimated by quantitative PCR. All the hepatitis-negative participants were vaccinated against HBV. The demographic and risk factors-related data were collected using the questionnaire. Statistical analysis (Chi-square test and bivariate regression analysis) was performed using SPSS software to explore any association between risk factors and hepatitis. Results indicated that 662/12912 participants (students = 478/10,948, employees = 184/1,964) tested positive for hepatitis. Among them, HCV was observed to be more prevalent than HBV among the study participants, employees, and students, and viral count was low in both HBV and HCV-infected participants. However, men were more affected than women. The studied risk factors represented higher frequency among hepatitis-positive participants relative to the hepatitis-negative participants. The Chi-square test revealed that students' gender, history of hepatitis in the family and relatives, dental treatment, sharing cosmetics and shaving blades were significant (p > 0.005) risk factors of hepatitis while in the employees group surgery and age were significant. Moreover, the reused of syringes was found to be associated with hepatitis in both groups. The bivariate analysis helped to identify various new risk factors which were independently, either positively or negatively, associated with hepatitis. Our study enabled us to recognize different risk factors of hepatitis among the target population. The information thus generated can be usefully applied in planning hepatitis awareness, targeted screening, and effective control programs for other target populations. In general, this module can be further utilized for any other disease.

Bacteremia with Multi-drug Resistant Gram-negative Bacteria in Pediatrics and Its Correlation with COVID-19

Background: Bloodstream infection with multi-drug resistant (MDR) bacteria has been introduced as the main risk factor for in-hospital mortality in vulnerable children worldwide. COVID-19 can complicate the treatment process in patients with bacteremia; however, data about this co-infection in children are scant. Objectives: This was a study on the antimicrobial patterns of Gram-negative bacteria (GNB) isolated from blood samples of children with bacteremia and their correlation with COVID-19. Methods: In this cross-sectional study, blood samples of children with bacteremia were analyzed using BACTEC bottles. The bacterial isolates were characterized based on standard microbiology laboratory methods, and MDR strains were detected based on a standard protocol. Real-time PCR tests for COVID-19 were recorded from the patients’ hospital documents. Results: A total of 255 positive blood samples were detected in children with bacteremia. The bacterial isolates included Enterobacteriaceae spp. 43.5% (111/255), Pseudomonas spp. 33.7% (86/255), Acinetobacter spp. 21.6% (55/255), and Stenotrophomonas spp. 1.2% (3/255). Of 255 GNB, 86.66% (221/255) were MDR, and the frequency of MDR strains was as follows: Enterobacteriaceae spp. 91.8% (102/111), Pseudomonas spp. 77.9% (67/86), Acinetobacter spp. 89% (49/55), and Stenotrophomonas spp. 100% (3/3). Of 255 children with GNB-related bacteremia, COVID-19 infection was confirmed in 25.1% (64/255) of them. Nearly 93.7% (60/64) of these patients had both MDR bacteremia and COVID-19. The correlation was significant between MDR bacteremia and COVID-19 (P-value = 0.002). The death rate was 43.33% (26/60) among these children. Conclusions: The results of this study showed that MDR-GNB was the main cause of bacteremia in children. Our findings showed a notable risk of concomitant COVID-19 and GNB-related bacteremia in these patients.

Performance of a Novel Real-Time PCR-Based Assay for Rapid Monkeypox Virus Detection in Human Samples.

The ongoing epidemic of mpox, namely human monkeypox virus (MPXV) infection, requires rapid and reliable laboratory diagnosis. We report on the QIAstat-Dx viral vesicular panel PCR assay that allows the detection of (within 75 min) six vesicular disease-causing viruses, including MPXV. We analyzed 168 clinical samples, known to be positive (51 samples) or negative (117 samples) for MPXV clade II, obtained from patients at their mpox diagnosis or follow-up. QIAstat assay results were compared to those of a MPXV-specific reference PCR assay. The QIAstat assay detected MPXV (clade II) in 51 (100%) of 51 samples and did not detect MPXV in 117 (100%) of 117 samples, resulting in a positive or negative agreement of 100% (95% CI, 93.0-100) and 100% (95% CI, 96.8-100), respectively. Of the 20 patients diagnosed with mpox, 18 (90.0%) had at least a vesicular swab and 1 (5.0%) had only an oropharyngeal swab positive for MPXV. At mpox follow-ups, 2 (10.0%) of 20 patients had first-time positive whole blood samples. Thirteen MPXV-negative samples were positive for mpox-mimicking viruses. Our findings show the excellent performance of the QIAstat-Dx assay for MPXV detection in clinical samples. Further studies are needed before considering a large-scale application of the QIAstat-Dx assay.

Clinical characteristics of patients with KRAS mutation detected by liquid biopsy.

KRAS mutation positive lung cancer is known to be clinically characterized by older age, males, and smokers. It is reported to be more common in mucinous adenocarcinoma, but all reports are based on analysis of tissue samples. Recently, blood samples have become available for analysis, suggesting a low detection rate of circulating tumor DNA in histological types, especially mucinous adenocarcinoma. In this study, we investigated the clinical characteristics of KRAS mutation-positive cases in the analysis of blood specimens, as these remain unclear. The clinical background of patients with KRAS mutation among those who underwent next-generation sequencing (NGS) analysis using blood samples was evaluated. NGS analysis was performed on 214 blood samples. KRAS mutations were detected in blood samples in 33 cases (15.4%), of which 31 cases (14.5%) had a histological pathology diagnosis. Mucinous adenocarcinoma accounted for 28.6% of cases with positive blood and tissue specimens, 10.0% of cases with positive blood specimens only, and 57.1% of cases with positive tissue specimens only. Mucinous adenocarcinoma tended to be less common in cases with positive blood specimens. In KRAS-positive patients with lung metastasis only, only one nonmucinous adenocarcinoma had a positive blood sample, and the others all had mucinous adenocarcinomas with positive tissue samples only. The results showed that the detection rate of KRAS-positive lung cancers detected by blood and tissue samples differs, and that the detection rate of blood samples may be poor, especially in the case of mucinous adenocarcinoma with lung metastases only.