Hematological Trends in Severe Burn Patients: A Comprehensive Study for Prognosis and Clinical Insights.

Severe burn injuries pose diagnostic challenges, contributing to increased fatality rates with delayed diagnoses. This study aims to identify early risk factors and understand their impact on clinical outcomes by examining hematological dynamics in severe burn cases. The focus includes age-related patterns, Total Body Surface Area (TBSA) affected by burns, hospital stay duration, and changes in hematological markers during burn injuries. An analytical cross-sectional study at the Burn Care Centre involved 135 participants hospitalized between January 2018 and December 2021. Demographic data and hematological markers were recorded, with statistical analysis using IBM SPSS 25.0. Non-survivors exhibited a greater mean TBSA, shorter hospital stay, and an enhanced early immune response indicated by WBC count on the first day. Hematological markers, including HGB, RCC, and PLT, showed dynamic patterns over the study period. Marginal variations in platelet counts and intriguing patterns in RCC suggested potential consequences like disseminated intravascular coagulation. The study provides crucial insights into hematological responses to severe burn injuries. Early identification of risk factors, particularly age-related patterns and immune responses, informs clinicians about predicting outcomes and guiding therapeutic interventions. Despite limitations, this work underscores the need for further multi-center research to comprehensively understand the complex relationships between burn injuries, hematological responses, and clinical outcomes.

Domestic Cylinder Blasts: A Neglected Torpedo.

High-pressure cylinders are used to store liquefied petroleum gas (LPG). An instant and swift explosion of these cylinders can result in serious burn injuries. This current research was conducted to study the epidemiological characteristics of LPG-related burns to highlight a major public health issue. Analysis was conducted on patients with LPG-related burns over a span of 10 years admitted to our center between January 2011 and December 2020. The data recorded included demographic features, site of burn, %TBSA, associated injuries, and outcomes. The variable data were documented for every patient in a Microsoft Excel file and analyzed by IBM SPSS version 25.0. Over the span of 10 years, 678 patients were affected by LPG-related accidents. The peak incidence was seen in 2019 when there was a surge to 18.03%. The patient's age ranged from 1 to 79 years, with a median of 40.86 ± 15.27 years. Of the 678 patients, 52.50% were males and 47.50% were females. The majority (57.96%) of patients had a total BSA of >60% and 86.72% were diagnosed with inhalation injury. The majority of burns (84.66%) occurred at home. The mean hospital stay was 24.5 days. The total mortality rate was 59.58%. This study concludes that LPG cylinder blast is a preventable cause that can be minimized by making people aware of its safe use and by arranging awareness programs at every national level.

Detection and Phylogenetic Analysis of Extended-Spectrum ß-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage.

Extended-spectrum ß-lactamases (ESBL) give rise to resistance against penicillin and cephalosporin antibiotics in multiple bacterial species. The present study was conducted to map genetic determinants and related attributes of ESBL-producing bacteria in three wild aquatic bird species and chickens at the "Trimmu Barrage" in district Jhang, Punjab province, Pakistan. To study the prevalence of ESBL-producing bacteria, a total of 280 representative samples were collected from wild bird species; cattle egrets (Bubulcus ibis), little egrets (Egretta garzetta) and common teals (Anas crecca) as well as from indigenous chickens (Gallus gallus domesticus) originating from a local wet market. The isolates were confirmed as ESBL producers using a double disc synergy test (DDST) and bacterial species were identified using API-20E and 20NE strips. A polymerase chain reaction (PCR) was used to detect ESBL genetic determinants and for genus identification via 16S rRNA gene amplification. A phenotypic antimicrobial susceptibility test was performed for ESBL-producing isolates against 12 clinically relevant antibiotics using the Kirby-Bauer disk diffusion susceptibility test. A phylogenetic tree was constructed for the sequence data obtained in this study and comparative sequence data obtained from GenBank. The overall prevalence of ESBL-producing bacteria was 34.64% (97/280). The highest percentage (44.28%; 31/70) of ESBL-producing bacteria was recovered from chickens (Gallus gallus domesticus), followed by little egrets (Egretta garzetta) (41.43%; 29/70), common teal (Anas crecca) (28.57%; 20/70) and cattle egrets (Bubulcus ibis) (24.28%; 17/70). Five different ESBL-producing bacteria were identified biochemically and confirmed via 16S rRNA gene sequencing, which included Escherichia coli (72; 74.23%), Enterobacter cloacae (11; 11.34%), Klebsiella pneumoniae (8; 8.25%), Salmonella enterica (4; 4.12%) and Pseudomonas aeruginosa (2; 2.06%). Based on PCR, the frequency of obtained ESBL genes in 97 isolates was blaCTX-M (51.55%), blaTEM (20.62%), blaOXA (6.18%) and blaSHV (2.06%). In addition, gene combinations blaCTX-M + blaTEM, blaTEM + blaOXA and blaCTX-M + blaSHV were also detected in 16.49%, 2.06% and 1.03% of isolates, respectively. The ESBL gene variation was significant (p = 0.02) in different bacterial species while non-significant in relation to different bird species (p = 0.85). Phylogenetic analysis of amino acid sequence data confirmed the existence of CTX-M-15 and TEM betalactamases. The average susceptibility of the antibiotics panel used was lowest for both Klebsiella pneumoniae (62.5% ± 24.42) and Salmonella enterica (62.5% ± 31.08) as compared to Enterobacter cloacae (65.90% ± 21.62), Pseudomonas aeruginosa (70.83% ± 33.42) and Escherichia coli (73.83% ± 26.19). This study provides insight into the role of aquatic wild birds as reservoirs of ESBL-producing bacteria at Trimmu Barrage, Punjab, Pakistan. Hence, active bio-surveillance and environment preservation actions are necessitated to curb antimicrobial resistance.

Cross-Sectional Study for Detection and Risk Factor Analysis of ESBL-Producing Avian Pathogenic Escherichia coli Associated with Backyard Chickens in Pakistan.

The increasing incidence of extended-spectrum ß-lactamase (ESBL)-producing Escherichia (E.) coli in backyard chicken farming in Pakistan is of serious concern. This study aimed to assess the prevalence, antimicrobial resistance patterns and risk factors associated with ESBL avian pathogenic E. coli (APEC) isolated from backyard chickens in the Jhang district, Punjab, Pakistan. In total, 320 cloacal swabs were collected from four breeds of backyard chicken (Aseel, Golden, Misri and Necked Neck). ESBL E. coli were phenotypically identified using double disc synergy test (DDST) and corresponding genes were confirmed by multiplex polymerase chain reaction (mPCR). Out of the 320 samples, 164 (51.3%) were confirmed as E. coli, while 74 (45.1%) were characterized as ESBL E. coli. The frequency of isolation of ESBL E. coli was highest in Aseel chickens (35.1%). Of the 164 confirmed E. coli, 95.1%, 78.6%, 76.8%, 71.3%, 70.1%, 68.9%, 60.4% and 57.3% were resistant against tylosin, doxycycline, cefotaxime, enrofloxacin, colistin, trimethoprim/sulfamethoxazole, chloramphenicol and gentamicin, respectively. The ESBL gene types detected and their corresponding proportions were blaCTX-M (54.1 %, 40/74), blaTEM, (12.2%, 9/74) and co-existence (blaCTX-M and blaTEM) were shown in 33.8% (25/74). The blaCTX-M gene sequence showed homology to blaCTX-M-15 from clinical isolates. The mean multiple antibiotic resistance index (MARI) was found to be higher among ESBL E. coli (0.25) when compared to non-ESBL E. coli (0.17). Both free-range husbandry management system (p = 0.02, OR: 30.00, 95% CI = 1.47-611.79) and high antimicrobial usage in the last 6 months (p = 0.01, OR: 25.17, 95% CI = 1.81-348.71) were found significantly associated with isolation of ESBL-producing E. coli in the tested samples using binary logistic regression analysis. This study confirmed the potential of backyard chickens as a reservoir for ESBL E. coli in the Jhang district, Punjab, Pakistan.

Treadmill Versus Overground Gait Training in Patients with Lower Limb Burn Injury: A Comparative Study.

The primary goal of the rehabilitation services offered to patients with lower limb burns is to enhance gait function. Exercise on a treadmill and conventional overground gait training are the two most widespread gait-training methods. Numerous studies have been published in the literature that claim treadmill training helps people walk more easily. Therefore, it would be intriguing to investigate whether treadmill gait training affects the progress made by post-lower limb burn patients compared to traditional overground gait training. The goal of the current study was to compare the effectiveness of treadmill training and traditional physical therapy therapies for improving gait after lower limb burn injury. A comparative study was conducted between January to August 2022 at a burns care unit, in Islamabad. All the participants were informed about the study procedure and consent was taken before initiating the study. All the participants received the set standard of Burn Rehabilitation by the American Burn Association (ABA). The program consists of ROM exercises, mobilization, functional mobility, stretching, balance training, and resisted exercises for enhancing muscle power. Statistical analyses were performed using IBM SPSS 24.0 software. The study subjects selected were 30 burn patients, aged above 18 years, vitally stable, and had lower limb burn injuries with an average TBSA (total burn surface area) of 20-30%. The mean for the Experimental is 18.86 and for the control group is 21.13. Patients were randomly either assigned to an experimental group or a control group. The Experimental group had a lower Time Up and Go test score of 11.86 ± 3.58 (9.50-24.07) as compared to the control group's TUG score (of 12.78 ± 4.41 (6.30-24.07). Functional mobility significantly improved as the P < 0.05. The total mean scores calculated on the Tampa Kinesiophobia scale for the Experimental group were 35.66 ± 6.32 which was less compared with those of the control group 37.93 ± 6.36. It demonstrates that fear of movement was reduced for treadmill training participants. Comparison of pre-distance (350.9 ± 86.82), pre-velocity (62.78 ± 20.20), and post-distance (385.02 ± 32.29), post-velocity 40.48 ± 21.91 of the experimental group with the control group pre-distance (339.26 ± 98.17), pre-velocity (55.07 ± 22.63), post-distance (383.07 ± 36.914), and post-velocity (40.57 ± 21.95) showed significant changes in results. In contrast to traditional gait training, treadmill training significantly improves walking ability in burn patients with lower limb damage.

Reimagining safe lithium applications in the living environment and its impacts on human, animal, and plant system.

Lithium's (Li) ubiquitous distribution in the environment is a rising concern due to its rapid proliferation in the modern electronic industry. Li enigmatic entry into the terrestrial food chain raises many questions and uncertainties that may pose a grave threat to living biota. We examined the leverage existing published articles regarding advances in global Li resources, interplay with plants, and possible involvement with living organisms, especially humans and animals. Globally, Li concentration (<10-300 mg kg-1) is detected in agricultural soil, and their pollutant levels vary with space and time. High mobility of Li results in higher accumulation in plants, but the clear mechanisms and specific functions remain unknown. Our assessment reveals the causal relationship between Li level and biota health. For example, lower Li intake (<0.6 mM in serum) leads to mental disorders, while higher intake (>1.5 mM in serum) induces thyroid, stomach, kidney, and reproductive system dysfunctions in humans and animals. However, there is a serious knowledge gap regarding Li regulatory standards in environmental compartments, and mechanistic approaches to unveil its consequences are needed. Furthermore, aggressive efforts are required to define optimum levels of Li for the normal functioning of animals, plants, and humans. This review is designed to revitalize the current status of Li research and identify the key knowledge gaps to fight back against the mountainous challenges of Li during the recent digital revolution. Additionally, we propose pathways to overcome Li problems and develop a strategy for effective, safe, and acceptable applications.

An automated design pipeline for transparent facial orthoses: A clinical study.

STATEMENT OF PROBLEM: Transparent facial orthoses (TFOs) are commonly used for the treatment of craniomaxillofacial trauma and burns to prevent hypertrophic and keloid scarring. A TFO is typically customized to the patient's facial contours and relies on a precise fit to ensure good rehabilitative performance. A smart method of TFO design and manufacture is needed which does not require an experienced prosthetist, allowing for rapidly produced, well-fitting TFOs. Whether the rapid application reduces the final level of patient scarring is unclear. PURPOSE: The purpose of this clinical study was to determine whether a scalable, automated design-through-manufacture pipeline for patient specific TFO fabrication would be successful. MATERIAL AND METHODS: The automated pipeline received a 3-dimensional (3D) facial scan captured from a depth sensitive mobile phone camera. The scan was cleaned, aligned, and fit to a template mesh, with a known connectivity. The resultant fitted scan was passed into an automated design pipeline, outputting a 3D printable model of a custom TFO. The TFOs were fabricated with 3D printing and were both physically and digitally evaluated to test the fidelity of a digital fit testing system. RESULTS: A total of 10 individuals were scanned with 5 different scanning technologies (STs). All scans were passed through an automated fitting pipeline and categorized into 2 groups. Each ST was digitally fitted to a ground truth scan. In this manner, a Euclidean distance map was built to the actual facial geometry for each scan. Heatmaps of 3D Euclidean distances were made for all participant faces. CONCLUSIONS: The ability to automatically design and manufacture a custom fitted TFO using commercially available 3D scanning and 3D printing technology was successfully demonstrated. After considering equipment size and operational personnel requirements, vat polymerization (VP) technology was found to be the most promising route to TFO manufacture.

Seroprevalence of Sars-Cov-2 antibodies among eligible blood donors of Peshawar, Pakistan.

BACKGROUND AND OBJECTIVES: To determine the seroprevalence of SARS-CoV-2 antibodies and the associated risk factors among healthy blood donors from Peshawar Pakistan, during the second and third waves of the COVID-19 pandemic. METHODS: The study was conducted on 4047 healthy (with no history or symptoms of COVID-19) blood donors attending regional blood center Peshawar between Nov 2020 and June 2021. Demographic data was collected and donors were screened for the presence of anti-SARS-CoV-2 antibodies using electrochemiluminescence immunoassay (ECLIA). RESULTS: The mean age of the participants was 27.27±7.13 and the majority (99%) were males. Overall, 59% (2391/4047) of the blood donors were reactive for SARS-CoV-2 antibodies. An increasing trend in seropositivity was observed from 45.5% to 64.8% corresponding to the second and third wave of the pandemic in Pakistan. Logistic regression analysis revealed significantly higher odds of seropositivity among male donors compared to females. Similarly, in multivariable analysis, the odds ratio for seropositivity among blood types AB, A, and B were, 1.6, 1.4, and 1.3 (CI 95%) times higher compared to blood group O (P-value ≤0.0001). CONCLUSIONS: Seropositivity of SARS-CoV-2 antibodies among blood donors gradually increased during the second and third wave of the pandemic in Pakistan indicating a widespread prevalence of Covid-19 in the general population. Susceptibility to SARS-CoV-2 varies with ABO blood types, with blood group O associated with low risk of infection.

First Report from Afghanistan on the Prevalence of Blood-Borne Infections: A Retrospective Cross-Sectional Multicentre Study for an Epidemiological Assessment.

BACKGROUND: The transfusion of blood and blood components has a significant role in healthcare services. However, it remains a possible risk factor for blood-borne infections. The present study was conducted to assess the prevalence of serological markers of common blood-borne infections among the blood donor population of Afghanistan. METHODOLOGY: This was a cross-sectional study based on retrospectively collected data over a period of six years from 284 blood centres across 34 provinces of Afghanistan. Every blood donor's sample was tested by rapid immunoassays for the serological markers of blood-borne infections namely hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV), anti-human immunodeficiency virus 1/2 (anti-HIV1/2), and anti-Treponema pallidum (anti-TP). RESULTS: All blood donors during the study period were males. The majority of blood donations were from the family replacement category 56.93% (n = 544,568). The overall pooled prevalence of blood-borne infections was 4.36% with a comparatively higher percentage in family replacement donors 4.88%. The seropositivity for HBsAg, anti-HCV, anti-HIV1/2, and anti-TP was 2.95%, 0.81%, 0.04%, and 0.54%, respectively. CONCLUSION: Complete reliance on voluntary blood donors and screening with quality assured highly sensitive assay is recommended to ensure blood safety in the country.

Toward Mass Customization Through Additive Manufacturing: An Automated Design Pipeline for Respiratory Protective Equipment Validated Against 205 Faces.

Respiratory protective equipment (RPE) is traditionally designed through anthropometric sizing to enable mass production. However, this can lead to long-standing problems of low-compliance, severe skin trauma, and higher fit test failure rates among certain demographic groups, particularly females and non-white ethnic groups. Additive manufacturing could be a viable solution to produce custom-fitted RPE, but the manual design process is time-consuming, cost-prohibitive and unscalable for mass customization. This paper proposes an automated design pipeline which generates the computer-aided design models of custom-fit RPE from unprocessed three-dimensional (3D) facial scans. The pipeline successfully processed 197 of 205 facial scans with <2 min/scan. The average and maximum geometric error of the mask were 0.62 mm and 2.03 mm, respectively. No statistically significant differences in mask fit were found between male and female, Asian and White, White and Others, Healthy and Overweight, Overweight and Obese, Middle age, and Senior groups.

A Systematic Review and Meta-Analysis on the Epidemiology of Hepatitis B and Hepatitis C Virus among Beta-Thalassemia Major Patients in Pakistan.

Beta-thalassemia major patients are the leading consumers of blood transfusions in Pakistan and, therefore, have a greater risk of acquiring transfusion-transmitted infections, most notably hepatitis B and C virus (HBV and HCV). The present study includes a comprehensive review on the status of HBV and HCV in beta-thalassemia major patients in Pakistan. For this purpose, we examined original articles assessing the epidemiology of HBV and HCV in transfusion-dependent thalassemia patients. We searched 10 major subscription databases from January through February 2020, that is, Medline, PakMediNet, CINAHL, Scopus, PubMed, Web of Science, Embase, Science Direct, Google Scholar, and Directory of Open Access Journals. The World Health Organization resources were also explored for relevant reports. The search criteria included published articles up to December 31, 2019, with no language restrictions. Articles identified were introduced into the Endnote version X9 software and then screened for relevance and duplication. The results were stated as the pooled prevalence for the overall study and also for region-wise subgroups. A total of 33 studies conducted from 1995 to 2019 were included in the review. All 33 articles yielded information on HCV prevalence, while 19 of them provided information on HBV prevalence. The overall sample size was 8,554 that tested the prevalence of HCV in thalassemia patients. The sample size from the 19 studies that tested the prevalence of HBV was 6,184. The overall pooled prevalence of HBV was computed to be 4.13%, while the pooled prevalence of HCV was 29.79%. The majority of the studies were obtained from the Punjab Province (33.33%), followed by Khyber Pakhtunkhwa Province (24.24%). The total sample size of 33 studies was less than 10% of the total number of estimated thalassemic patients, that is, 100,000. Further studies or a national baseline survey are imperative to confirm the actual frequency of HBV and HCV in thalassemia patients across the country.

Seroprevalence of Transfusion-Transmitted Infections among Voluntary and Replacement Blood Donors at the Peshawar Regional Blood Centre, Khyber Pakhtunkhwa, Pakistan.

Introduction Blood transfusion is linked to several risks, most notably the transmission of transfusion-transmitted infections (TTIs), including hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), syphilis, and malaria. The risk posed by these blood-borne infectious agents is high in developing countries, including Pakistan. This fact stresses the need for regular surveillance of TTIs. Therefore, the present study was undertaken to assess the seroprevalence of TTIs at a regional blood center. Material and Methods This was a retrospective 4-year descriptive study undertaken at the Regional Blood Centre in Peshawar, Khyber Pakhtunkhwa Province of Pakistan, on the blood donor data from June 2016 to May 2020. A total of 41,817 donors donated blood during the study period and were screened for HBV, HCV, HIV, syphilis, and malaria. To ensure donor privacy, donors were identified via codes and no personal information was available. The data were extracted from the ZAAVIA blood transfusion information system database. Results The study included a total of 41,817 donors-41,493 (99.22%) males and 324 (0.78%) females. Of them, 22,343 (53.43%) were voluntary donors while 19,474 (46.57%) were replacement donors. An overall TTI prevalence rate of 4.61% was found. The TTI prevalence rate in voluntary donors was 3.90% while 5.42% in replacement donors. The overall prevalence of HBV, HCV, HIV, syphilis, and malaria was 1.95, 1.38, 0.23, 0.91, and 0.14%, respectively. Conclusion The current study documented a high prevalence (1,929 out of 41,817, 4.61%) of TTIs, especially in replacement donors (1,057 out of 19,474, 5.42%), and low participation of female donors. The recommendations include the promotion of voluntary blood donors, enrolment of female blood donors, and screening of donated blood through highly sensitive screening assay (i.e., nucleic acid testing).

Molecular and genetic characterization of hepatitis B virus among multitransfused thalassaemia patients in Islamabad, Pakistan.

BACKGROUND: Hepatitis B virus (HBV) is the aetiological agent of transfusion-transmitted hepatitis globally. Beta thalassaemia major individuals are at greater risk of contracting HBV infection due to multiple blood transfusions required for the medical management of these patients. Based on HBV genetic variability, it is divided into 10 genotypes. The determination of HBV genotypes has significant implications for clinical management and treatment regimens. AIM: This study was performed to assess the HBV epidemiology and circulating genotypes in multi-transfused ß-thalassemia major patients with the aim to be considered while formulating the treatment pattern taking into account particular needs of thalassaemia patients. MATERIALS AND METHODS: This study was performed from September 2018 to June 2019, at the Department of Pathology and Transfusion Medicine, Shaheed Zulfiqar Ali Bhutto (SZAB) Medical University, Islamabad. A total of 2,260 thalassaemia patients were enrolled in the study. The study was endorsed by the Ethics Committee of the SZAB Medical University, Islamabad. The samples were serologically screened for HBsAg on the LIAISON® XL Murex HBsAg Quant assay (DiaSorin S.p.A., Italy) a chemiluminescence based immunoassay (CLIA). HBV quantitative PCR kit was used to measure the HBV DNA in serum samples. The HBV genotypes were determined using universal primers targeting the P1 and S1 region amplification. RESULTS: Of 2,260 thalassaemia patients, 64.6% were males while 35.4% were females. The HBsAg was identified in 98 individuals (4.33%). The PCR analysis was done for these 98 patients and in this cohort, genotype D was 59.18% (n = 58), genotype A was 21.42% (n = 21) while genotype C was 19.38% (n = 19). CONCLUSION: The determination of HBV genotypes in the multi-transfused patients is key to the effective management of chronic HBV patients as the severity and course of the disease is dependent on a specific type of genotypes. Quality assured screening of donated blood will prevent the incidence of HBV in thalassaemia patients.

Seroprevalence and Molecular Identification of Brucella spp. in Bovines in Pakistan-Investigating Association With Risk Factors Using Machine Learning.

Bovine brucellosis is a global zoonosis of public health importance. It is an endemic disease in many developing countries including Pakistan. This study aimed to estimate the seroprevalence and molecular detection of bovine brucellosis and to assess the association of potential risk factors with test results. A total of 176 milk and 402 serum samples were collected from cattle and buffaloes in three districts of upper Punjab, Pakistan. Milk samples were investigated using milk ring test (MRT), while sera were tested by Rose-Bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (i-ELISA). Real-time PCR was used for detection of Brucella DNA in investigated samples. Anti-Brucella antibodies were detected in 37 (21.02%) bovine milk samples using MRT and in 66 (16.4%) and 71 (17.7%) bovine sera using RBPT and i-ELISA, respectively. Real-time PCR detected Brucella DNA in 31 (7.71%) from a total of 402 bovine sera and identified as Brucella abortus. Seroprevalence and molecular identification of bovine brucellosis varied in some regions in Pakistan. With the use of machine learning, the association of test results with risk factors including age, animal species/type, herd size, history of abortion, pregnancy status, lactation status, and geographical location was analyzed. Machine learning confirmed a real observation that lactation status was found to be the highest significant factor, while abortion, age, and pregnancy came second in terms of significance. To the authors' best knowledge, this is the first time to use machine learning to assess brucellosis in Pakistan; this is a model that can be applied for other developing countries in the future. The development of control strategies for bovine brucellosis through the implementation of uninterrupted surveillance and interactive extension programs in Pakistan is highly recommended.

Genetic Diversity of Human Immunodeficiency Virus Type 1 in Asymptomatic Blood Donors in Islamabad, Pakistan.

Objective The serological testing of human immunodeficiency virus (HIV) is mandatory under the blood safety legislation of Pakistan; hence, data exist on the prevalence of HIV in blood donors. However, little is known about the molecular epidemiology of HIV in the blood donor population. Therefore, the current study was designed to study the genetic diversity of HIV-1 infection in a population of apparently healthy treatment-naive blood donors in Islamabad, Pakistan. Material and Methods A total of 85,736 blood donors were tested for HIV by the chemiluminescence immunoassay. All positive donor samples were analyzed for the presence of various HIV genotypes (types and subtypes). Viral ribonucleic acid was extracted from blood samples of HIV positive donors and reverse transcribed into complementary deoxyribonucleic acid (cDNA). The cDNA of all positive donors was then analyzed for the presence of various HIV genotypes (types and subtypes) by employing subtype-specific primers in a nested polymerase chain reaction. The amplified products were run on ethidium bromide-stained 2% agarose gel and visualized using a ultraviolet transilluminator. A particular subtype was assigned to a sample if the subtype-specific reaction made a band 20% highly intense compared with the band made by the subtype-independent reaction. Results A total of 85,736 blood donors were screened for the presence of antibodies to HIV. Out of them, 114 were initially found reactive for HIV. The repeat testing resulted in 112 (0.13%) positive donors, 95% confidence interval 0.0014 (0.0011-0.0018). These 112 samples were analyzed for molecular typing of HIV-1. The predominant HIV-1 subtype was A ( n = 101) (90.1%) followed by subtype B ( n = 11) (9.9%). Conclusion These findings are key to understand the diversified HIV epidemic at the molecular level and should assist public health workers in implementing measures to lessen the further dissemination of these viruses in the country.

Molecular epidemiology of hepatitis B virus genotypes in blood donors in Islamabad, Pakistan.

BACKGROUND: Hepatitis B virus (HBV) is a major causative agent of early, severe and prolonged liver infection that subsequently leads to cirrhosis of liver and hepatocellular carcinoma. The aim of this study was to evaluate the molecular epidemiology of hepatitis B virus (HBV) genotypes and comparison of serological assay performance versus polymerase chain reaction (PCR) in HBV screening. METHODS: Blood samples of 8517 healthy blood donors were collected during the period of January to June 2017 from Blood Bank of Shaheed Zulfiqar Ali Bhutto Medical University, Islamabad. Samples were screened for HBsAg assay using technique of chemiluminescence immunoassay. PCR of positive samples was carried out using already reported genotype-specific primers by Naito et al. (2001). The results were confirmed by visualizing genotype bands. RESULTS: The study confirmed the presence of HBV in 2.5% of blood donors, and PCR confirmed the presence of HBV-DNA in 92 samples. The genotyping was done by PCR using type-specific primer sequences. PCR was dogged to check six genotypes, i.e., A, B, C, D, E, and F. The results of this study show high levels of Genotype D is this region, i.e., 52.17% with less dominating Genotype C, which is 16.30% with decreasing ratio of Genotype E (14.13%), Genotype A and B (9.78%), and mixed D + E (2.17%). The presence of coinfection is found at lowest rate. Due to the high percentage of HBV/D, it is concluded that D genotype is common in our population. CONCLUSION: The most prevalent HBV genotype in ICT region was genotype D, which is responsible for liver cirrhosis and hepatocellular carcinoma. Efficacy of drugs varies with variation in genotypes of hepatitis B virus and also with geographical distribution.

Evaluation of screening effectiveness of hepatitis B surface antigen and anti-HCV rapid test kits in Pakistan.

OBJECTIVE: The current study was conducted to evaluate the performance and screening effectiveness of commercially available rapid screening kits in comparison with chemiluminescence immunoassay (CLIA) and polymerase chain reaction (PCR). MATERIALS AND METHODS: This single-center, cross-sectional study was conducted at the Department of Pathology and Blood Transfusion Services, Shaheed Zulfiqar Ali Bhutto Medical University, PIMS, Islamabad, from January to April 2019. A total of 10 commercially available immunochromatographic test (ICT) devices and one CLIA kit (LIAISON XL) were tested for their sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy using 100 positive and 100 negative samples each for HBV and HCV, respectively. RESULTS: The sensitivities and specificities of ICT kits for hepatitis B surface antigen were 65% and 70% (Hightop), 67% and 85% (RightSign), 62% and 73% (Wondfo), 70% and 80% (Accu-Chek), 68% and 77% (Fastep), 73% and 85% (Abon), 77% and 83% (ImmuMed), 80% and 90% (Insta-Answer), 67% and 81% (BioCheck), and 72% and 83% CTK Biotech, respectively. Similarly, the sensitivities and specificities of different ICT kits for HCV were 69% and 80% (Hightop), 76% and 83% (RightSign), 69% and 81% (Wondfo), 78% and 79% (Accu-Check), 68% and 68% (Fastep), 63% and 73% (Abon), 71% and 70% (ImmuMed), 79% and 68% (Insta-Answer), 62% and 66% (BioChek), and 69% and 78% CTK Biotech, respectively. The sensitivity and specificity of Diasorin Liaison Murex assay for both HBV and HCV were found to be 100% when compared with PCR. The PPV, NPV and Accuracy were determined accordingly. CONCLUSION: Rapid testing ICT devices for both HBV and HCV available in Pakistan were found to have a variable degree of sensitivity and specificity when compared with CLIA and PCR. Comparatively expensive but quality methods are more reliable as compared to rapid devices.