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 July and 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.

Investigation of mechanical and thermal behavior of fiber-reinforced silica xerogel composites.

Silica aerogels or xerogels are renowned dried gels with low density, high surface area, higher porosity, and better thermal stability which makes it suitable for aerospace, light weight structures, thermal insulation, and hydrophobic coatings. But brittle behaviour, low mechanical strength, and high manufacturing cost restrict its usage. Recently, the addition of various fibres like glass or carbon fiber is one of the best reinforcement methods to minimize the brittle behaviour. Supercritical drying technique usually used to develop aerogel that is expensive and difficult to produce in bulk quantities. Higher cost obstacle can be tackled by applying ambient pressure drying technique to develop xerogel. But researcher observed cracks in samples prepared through the ambient pressure drying technique is still a major shortcoming. The aim of this study is to systematically analyze the influence of silica gel fiber reinforcement on silica xerogels, encompassing morphology, mechanics, thermal behaviour, compression test, and thermogravimetric characteristics. The research used a low-cost precursor named Tetraethyl orthosilicate to synthesize low-cost composite Silica xerogel and glass and carbon fiber added to provide strength and flexibility to the overall composite. Silica gel works as binder in strengthening the xerogel network. The investigation employs scanning electron microscopy (SEM) to examine the morphology of the composites, Fourier Transform Infrared (FTIR) analysis to affirm hydrophobic characteristics, compression tests to assess mechanical strength, and thermogravimetric tests to study weight loss under different conditions. SEM results reveals that glass fibers exhibit lower adhesion to the xerogel network compared to carbon fibers. FTIR analysis confirms the hydrophobicity of the composite silica xerogel. Compression tests showed that, under a 48% strain rate, the carbon fiber composite demonstrates superior compressive stress endurance. Thermogravimetric tests revealed a 1% lower weight loss for the carbon fiber composite compared to the glass fiber composite. This work concludes that glass and carbon fiber together with silica gel particles successfully facilitated in developing flexible, less costly, hydrophobic, and crack-free silica xerogel composites by APD. These advancements have the potential to drive innovations in material science and technology across diverse industries.

Genotypic analysis of hepatitis E virus (HEV) from sporadic symptomatic cases in Pakistan.

Pakistan is the fifth most populous nation in the world and faces several challenges, including devastating floods, sub-optimal sanitary conditions, clustered accommodations, and unregulated cross-border movements. These drastic population shifts make it vulnerable to the efficient spread of the Hepatitis E virus (HEV). The current study analyzed the genotypic characteristics and variants of the Hepatitis E virus circulating in the population of Pakistan. A total of 75 ELISA-IgM positive samples were collected from three metropolitan cities: Lahore, Peshawar, and Karachi, and subjected to viral RNA extraction. The amplification of the HEV RNA-dependent RNA polymerase (RdRp) region was done using Nested PCR and degenerate primers. Out of the total, 40% of the samples were positive for HEV RNA. Sequencing and phylogenetic analysis identified the new HEV isolates as Subtype 1 g, a subtype within an existing HEV genotype 1. This shift warrants investigation into its impact on clinical manifestation and disease severity. Importantly, this study marks the first HEV subtype analysis in Pakistan, contributing valuable insights into subtype diversity and prevalence in the region.

Genomic characterization of the first mpox and varicella-zoster co-infection in Pakistan (2023) through next-generation sequencing.

The global mpox outbreak spanning 2022-2023 has affected numerous countries worldwide. In this study, we present the first report on the detection, whole-genome sequence, and coinfection of the mpox virus and varicella zoster virus (VZV) from Pakistan. During April-May 2023, samples from 20 suspected cases of mpox were tested at the National Institutes of Health, Islamabad among which 4 tested positive. All four cases had a travel history of Saudi Arabia. All the suspected samples were processed by using a Zymo research kit for DNA extraction, followed by qRT-PCR amplification by using a DaAn Gene detection kit for the mpox virus. Further, two of the positive samples with a low Ct value (<20) were subjected to whole-genome sequencing using a metagenomic approach on the iSeq (Illumina) platform. The sequencing results revealed Clade IIb and genotype A.2.1 of MPXV, which clustered with viruses from Slovenia and the UK in July and June 2022, respectively. Our analysis identified two novel nonsynonymous substitutions in mpox virus, namely V98I in OPG046 and P600S in OPG109. Furthermore, we successfully retrieved the complete genome of VZV from the same sample, belonging to Clade 5. This study represents the first positive case of MPXV in Pakistan and the coinfection of mpox and VZV by using a metagenome approach providing insights into their complete genomes. Our results highlight the importance of surveillance at the point of entries, strengthening lab capacities including next-generation sequencing, and using differential diagnosis for timely and accurate detection of mpox cases.

Clinical utility and diagnostic value of tumor-educated platelets in lung cancer: a systematic review and meta-analysis.

Background: The review addresses the knowledge gap concerning the diagnostic value and clinical utility of tumor-educated platelets (TEPs) in adult patients with lung cancer. Methods: We searched twelve databases: PubMed, CENTRAL, EMBASE, CINAHL, MEDLINE, Scopus, ProQuest, MedRxiv, BioRxiv, SSRN, Clinicaltrials.gov, and CNKI up to 24 March 2023, to include any diagnostic study regarding TEPs and LC. TEPs diagnostic value was evaluated from pooled sensitivity and specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and the area under the curve (AUC). QUADAS 2 was used to assess the risk of bias. Heterogeneity analysis was assessed using the receiver operating characteristic (ROC) plane, Galbraith plot, bivariate boxplot, sensitivity analysis, and meta-regression. TEPs clinical utility was evaluated from Fagan's nomogram. Results: 44 reports from 10 studies, including 7,858 events and 6,632 controls, were analyzed. The pooled sensitivity, specificity, PLR, NLR, and DOR were 0.80 (95% CI 0.79-0.80), 0.69 (95% CI 0.69-0.70), 2.92 (95% CI 2.50-3.41), 0.26 (95% CI 0.21-0.32), and 12.1 (95% CI 8.61-16.76), respectively. In addition, the AUC of the Summary ROC curve was 0.85 (95% CI: 0.81-0.88). The overall risk of bias was low. Heterogeneity may result from cancer stage, cancer control, measuring equipment, and RNA types across studies. There was no apparent publication bias (p=0.29) with significant positive (79%) and negative (22%) post-test probability, according to Deeks funnel plot asymmetry test and Fagan's nomogram. Conclusion: TEPs could be a moderately effective candidate biomarker for LC diagnosis.

A short review on the role of thyroxine in fast wound healing and tissue regeneration.

Wound healing is a multiplex interaction process that involves extracellular matrix, blood vessels, proteases, cytokines, and chemokine. So far, a number of studies have been performed to understand the basis of the wound-healing process and multiple wound-healing products have been designed. However, significant morbidity and mortality incidents still occurred due to poor wound healing. Thus, there is a dire need to understand the effects of topical applications of various therapeutic options that lead to fast wound healing. Thyroxine is one great panacea for wound healing that has been vigorously mooted throughout the years but a conclusive result regarding its effectiveness is still not achieved. This review is intended to find a rational basis for its positive role in wound healing. To accomplish the objective, this review highlights the different aspects of thyroxine's role in wound healing like keratin synthesis, skin thickening, and pro-angiogenesis, the basis of controversy on its wound healing ability and its potential to be used as a wound healing agent. This study will be helpful for researchers and surgeons to assess the importance of thyroxine as a candidate to comprehensively research to develop a potent, effective, and affordable wound healing drug.

Synthesis and applications of MXene-based composites: a review.

Recently, there has been considerable interest in a new family of transition metal carbides, carbonitrides, and nitrides referred to as MXenes (Ti3C2Tx) due to the variety of their elemental compositions and surface terminations that exhibit many fascinating physical and chemical properties. As a result of their easy formability, MXenes may be combined with other materials, such as polymers, oxides, and carbon nanotubes, which can be used to tune their properties for various applications. As is widely known, MXenes and MXene-based composites have gained considerable prominence as electrode materials in the energy storage field. In addition to their high conductivity, reducibility, and biocompatibility, they have also demonstrated outstanding potential for applications related to the environment, including electro/photocatalytic water splitting, photocatalytic carbon dioxide reduction, water purification, and sensors. This review discusses MXene-based composite used in anode materials, while the electrochemical performance of MXene-based anodes for Li-based batteries (LiBs) is discussed in addition to key findings, operating processes, and factors influencing electrochemical performance.

Frequency and survival of Covid associated mucormycosis patients at tertiary care hospitals in Pakistan: A retrospective observational study.

Background: Mucormycosis is a serious fungal infection associated with uncontrolled diabetes and immunocompromised patients. This angioinvasive infection emerged as a post-covid complication worldwide especially in developing countries. Due to the common socio-demographic status of South Asian countries, we expected a surge in mucormycosis cases in Pakistan. This study aims to observe the frequency and survival of Covid associated mucormycosis patients at tertiary care hospitals in Pakistan during the third wave of Covid-19 in 2021. Materials and Methods: In this retrospective study, we collected the data of clinically and histopathologically confirmed cases of rhino-occipito-cerebral mucormycosis from three tertiary care hospitals of Lahore. These cases were analysed for history of Covid-19 and other associated comorbidities using SPSS28. History of steroid medication was also taken. Data were retrieved from May to July 2021 after the approval from the ethical review board. Results: Out of the total 43 reported patients of mucormycosis in the set time frame only 22 cases had a history of Covid-19. The mean age was 50 ± 13.27 years with slight male predilection (60%). Diabetes mellitus was the most common comorbidity (88.4%) and all the patients with covid associated mucormycosis (CAM) had taken corticosteroid regimen for covid management (p < 0.0001). The survival of the patient was not significantly different between CAM and non-CAM patients of Mucormycosis (p = 0.747). Conclusion: Covid-19 and mucormycosis make a lethal duo against the weakened health system of Pakistan. This problem can be prevented by avoiding nonjudicial use of corticosteroids and proper diabetes control program following Covid-19 infection. Furthermore, large-scale epidemiological studies should be carried out to evaluate the true burden of Mucormycosis in the population.

Breakthrough cases of Omicron and Delta variants of SARS-CoV-2 during the fifth wave in Pakistan.

COVID-19 pandemic has severely affected Pakistan with 1,557,134 cases as of August 4, 2022. However, the data regarding breakthrough infections in Pakistan is scant. Hence, the objective was to analyze SARS-CoV-2 breakthrough infections with respect to vaccines and variants during the fifth wave in Pakistan. Therefore, the Department of Virology (NIH, Pakistan) genotyped 2,467 randomly selected individuals between November 2021 and February 2022 using the SNPsig® SARS-CoV-2 (EscapePLEX) kit (PrimerDesign, UK). P681R and K417N mutations were used to distinguish delta and omicron. Data on the patient's age, gender, date of collection, variant, and vaccination status were analyzed using Statistical Package for Social Sciences (SPSS) software. Among 2,467 genotyped samples, Omicron was detected in 58.6% (n = 1445), Delta in 40.4% (n = 998) and undetermined/wildtype variant in 24 samples. The vaccination status of omicron-positive patients showed (49.7%; n = 718/1445) and Delta-positive patients (39.67%; n = 396/998) to be fully vaccinated. Of note, a high percentage 85% of breakthrough cases (n = 947) were identified among fully vaccinated individuals (n = 1114). Among them, 85.9% (n = 617/718) belonged to omicron and 83.3% (n = 330/396) to delta. Moreover, 76.7% (n = 855) of vaccinated individuals (n = 1114) received Sinopharm (n = 432) and Sinovac (n = 423) vaccines. The majority of breakthrough subjects who contracted Omicron were vaccinated with Sinopharm (93.0%, n = 256) and delta with Cansino (100%, n = 44). Individuals vaccinated with Sinovac showed the most frequent breakthrough cases for both Omicron and Delta variant between the 4th and 6th months (n = 278) after primary vaccination as compared to the 7th to 9th months (n = 24) category. While in case of Sinopharm, maximum breakthrough cases occurred between 7th to 9th months (n = 234) as compared to the 4th to 6th months (n = 120) after primary vaccination. Omicron and Delta breakthrough cases in men (n = 364 and 193) are more frequently seen than women (n = 253 and 138) respectively and breakthrough majority cases (n = 392) occurred in individuals aged 18-33 years. Breakthrough cases limiting monitoring in Pakistan impose a substantial constraint on policymakers' ability to take timely effective decisions. Since the current study consists of only a 2,467-genotyped sample, comprehensive data should be analyzed.

Genomic diversity of SARS-CoV-2 in Pakistan during the fourth wave of pandemic.

The emergence of different variants of concern of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in upsurges of coronavirus disease 2019 (COVID-19) cases around the globe. Pakistan faced the fourth wave of COVID-19 from July to August 2021 with 314,786 cases. To understand the genomic diversity of circulating SARS-CoV-2 strains during the fourth wave of the pandemic in Pakistan, this study was conducted. The samples from 140 COVID-19-positive patients were subjected to whole-genome sequencing using the iSeq Sequencer by Illumina. The results showed that 97% (n = 136) of isolates belonged to the delta variant while three isolates belonged to alpha and only one isolate belonged to the beta variant. Among delta variant cases, 20.5% (n = 28) isolates were showing B.1.617.2 while 23.5% (n = 25), 17.59% (n = 19), 14.81% (n = 16), and 13.89% (n = 15) of isolates were showing AY.108, AY.43 AY.127, and AY.125 lineages, respectively. Islamabad was found to be the most affected city with 65% (n = 89) of delta variant cases, followed by Karachi (17%, n = 23), and Rawalpindi (10%, n = 14). Apart from the characteristic spike mutations (T19R, L452R, T478K, P681R, and D950N) of the delta variant, the sublineages exhibited other spike mutations as E156del, G142D, T95I, A222V, G446V, K529N, N532S, Q613H, and V483A. The phylogenetic analysis revealed the introductions from Singapore, the United Kingdom, and Germany. This study highlights the circulation of delta variants (B.1.617.2 and sublineages) during the fourth wave of pandemic in Pakistan.

Genomic surveillance reveals the detection of SARS-CoV-2 delta, beta, and gamma VOCs during the third wave in Pakistan.

SARS-CoV-2 variants of concern (VOCs) have emerged worldwide and gained significant importance due to their high transmissibility and global spread, thus meriting close monitoring. In Pakistan, limited information is available on circulation of these variants as the alpha variant has been reported the main circulating lineage. The current study was designed to detect and explore the genomic diversity of SARS-CoV-2 lineages circulating during the third wave of the pandemic in the indigenous population. From May 01 to June 09, 2021, a total of 16 689 samples were tested using TaqPath™ COVID-19 kit for the presence of SARS-CoV-2. Overall, 2562 samples (15.4%) were COVID-19 positive. Out of these positive samples, 2124 (12.7%) did not show the spike gene amplification (spike gene target failure ([SGTF]), whereas 438 (2.6%) showed spike gene amplification (non-SGTF). A subset (n = 58/438) of non-SGTF samples were randomly selected for whole-genome sequencing. Among VOCs, 45% (n = 26/58) were delta, 46% (n = 27/58) were beta, and one was gamma variant. The delta variant cases were reported mainly from Islamabad (n = 15; 58%) followed by Rawalpindi and Azad Kashmir (n = 1; 4% each). Beta variant cases originated mainly from Karachi (n = 8; 30%) and Islamabad (n = 11; 41%) and the gamma variant case was reported in a traveler from Italy. The delta, beta, and gamma variants possessed lineage-specific spike mutations. Notably, two rare mutations (E484Q and L5F) were found in the delta variant. Furthermore, in the beta variant, two significant rare non-synonymous spike mutations (A879S and K444R) were also reported. High prevalence of beta and delta variants in local population may increase the number of cases in the near future and provides an early warning to national health authorities to take timely decisions and devise suitable interventions to contain a possible fourth wave.

Direct growth of m-BiVO4@carbon fibers for highly efficient and recyclable photocatalytic and antibacterial applications.

Owing to photocatalytic and antibacterial properties, bismuth based oxides has drawn much attention in recent past. However, non-recyclability of these oxides has restricted their practical applications. In present work, a novel nanostructured composite monoclinic bismuth vanadate@ activated carbon fibers (BiVO4@ACF) photocatalyst was efficaciously synthesized using a solvothermal method and characterized by X-ray diffraction (XRD), scanning electron microscope (SEM) and Bruner-Emmett-Teller (BET). The specific surface area, phase composition, microstructure, binding and photocatalytic activity of BiVO4@ACF pose great dependence on solvent nature and chelating agents utilized for synthesis. The photocatalytic and antibacterial potential of this composite was evaluated and optimized by using a model pollutant, Reactive Rhodamine Blue (RhB) and pathogenic microbes (Escherichia coli and Staphylococcus aureus). The composite possesses enhanced photocatalytic and antibacterial activity and was reutilized for three rounds of respective reaction without any loss of activity and structure as evident from SEM and XRD results. The photocatalytic mechanism of photodegradation of dye and bactericidal properties of samples under visible light irradiation was determined by scavenger and photoluminescence (PL) spectroscopy. The enhanced photocatalytic and antibacterial activity, chemical stability and most importantly good recyclability of BiVO4@ACFs highlight the potential application of this composite in water purification and other biological applications.

Trichosporon Species and Fusarium Species as a Cause of Empyema Thoracis in a Diabetic Patient.

Of late, fungal infections are increasingly being recognized in diabetic patients. Here we present a case of polymicrobial fungal empyema due to Trichosporon species and Fusarium species developed after community-acquired pneumonia in a diabetic patient. Trichosporon species are basidiomycetous yeast and Fusarium species are soil saprophytes with a worldwide distribution. Fungal empyema cases are rare and are mostly caused by Aspergillus and Candida species. Polymicrobial fungal empyema with Trichosporon species and Fusarium species has not been reported previously. Our patient was successfully treated with antifungal therapy. This case highlights that fungal empyema should be considered in diabetic patients especially if they are not responding to antibiotics.

The Dilemma of Renal Involvement in COVID-19: A Systematic Review.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), now known as coronavirus disease 2019 (COVID-19), has posed a serious threat to global health since December 2019. It has spread worldwide and is consuming healthcare resources rapidly. Published literature suggests that people with advanced age and comorbidities are affected more severely. It is crucial to uncover the underlying pathogenesis of acute kidney insufficiency in COVID-19 patients to understand better the reasoning behind the grave outcomes in these patients. In this review, we have included articles stating the prevalence and specific mortality rates of COVID-19 patients with acute kidney insufficiency. Our study included 1098 COVID-19 positive patients, of whom 66 (6%) developed acute kidney insufficiency and 62 patients died, showing a mortality rate of 94%. Patients with acute kidney insufficiency showed a more severe disease course, and these patients ended up more in intensive care units. Particular attention should be paid to those with already established kidney disease, such as chronic kidney disease, or renal transplant recipients, as these patients are already on immunosuppressive therapy. Our review focuses on the prevalence of acute kidney insufficiency in COVID-19 patients and mortality rates in this subset of patients.

Z-scheme 2D-m-BiVO4 networks decorated by a g-CN nanosheet heterostructured photocatalyst with an excellent response to visible light.

For economical water splitting and degradation of toxic organic dyes, the development of inexpensive, efficient, and stable photocatalysts capable of harvesting visible light is essential. In this study, we designed a model system by grafting graphitic carbon nitride (g-C3N4) (g-CN) nanosheets on the surface of 2D monoclinic bismuth vanadate (m-BiVO4) nanoplates by a simple hydrothermal method. This as-synthesized photocatalyst has well-dispersed g-CN nanosheets on the surface of the nanoplates of m-BiVO4, thus forming a heterojunction with a high specific surface area. The degradation rate for bromophenol blue (BPB) shown by BiVO4/g-CN is 96% and that for methylene blue (MB) is 98% within 1 h and 25 min, respectively. The 2D BiVO4/g-CN heterostructure system also shows outstanding durability and retains up to ∼95% degradation efficiency for the MB dye even after eight consecutive cycles; the degradation efficiency for BPB does not change too much after eight consecutive cycles as well. The enhanced photocatalytic activities of BiVO4/g-CN are attributed to the larger surface area, larger number of surface active sites, fast charge transfer and improved separation of photogenerated charge carriers. We proposed a mechanism for the improved photocatalytic performance of the Z-scheme photocatalytic system. The present work gives a good example for the development of a novel Z-scheme heterojunction with good stability and high photocatalytic activity for toxic organic dye degradation and water splitting applications.

Mo-doped ZnO nanoflakes on Ni-foam for asymmetric supercapacitor applications.

A single-step hydrothermal route for synthesizing molybdenum doped zinc oxide nanoflakes was employed to accomplish superior electrochemical characteristics, such as a specific capacitance of 2296 F g-1 at current density of 1 A g-1 and negligible loss in specific capacitance of 0.01025 F g-1 after each charge-discharge cycle (up to 8000 cycles). An assembled asymmetric supercapacitor (Mo:ZnO@NF//AC@NF) also exhibited a maximum energy density and power density of 39.06 W h/kg and 7425 W kg-1, respectively. Furthermore, it demonstrated a specific capacitance of 123 F g-1 at 1 A g-1 and retained about 75.6% of its initial capacitance after 8000 cycles. These superior electrochemical characteristics indicate the potential of this supercapacitor for next-generation energy storage devices.

An Insight into the Anaerobic Co-digestion of Municipal Solid Waste and Food Waste: Influence of Co-substrate Mixture Ratio and Substrate to Inoculum Ratio on Biogas Production.

The unbalanced nutrients of municipal solid waste (MSW), particularly high carbon contents, were regarded as a major limiting factor to anaerobic digestion process. In this study, the addition of MSW in food waste (FW) feedstock to have a balanced C/N ratio was studied. Different co-substrate mixtures with C/N ratio of 20 to 40 were subjected to anaerobic batch experiment at lab scale, under mesophilic conditions. The biogas production decreased with the increase in C/N ratio due to insufficient availability of organic nitrogen for anaerobic microbial growth. Specific biogas and methane yields were observed to be 827 and 474.44 mL g-1VS, respectively, with volatile solids (VS) reduction rate of 88%, at C/N ratio of 20. Furthermore, the effect of the substrate to inoculum (S/I) ratio on digester performance was also studied. The biogas production decreased with the increase in S/I ratio due to the formation of more volatile fatty acids (VFAs) which led to decrease in pH and accumulated unionized ammonia-N. Specific biogas and methane yields were recorded to be 655 and 410.20 mL g-1VS, with 64% rate of biodegradability at S/I ratio of 0.5. Kinetics and statistics study showed that the higher S/I ratio could lead to VFA accumulation and result in low methane yield.

Diagnostically untypable hepatitis C virus variants: it is time to resolve the problem.

Pakistan is a low income country with more than 10 million hepatitis C virus (HCV) infections and the burden is on continuous raise. Accurate viral genotyping is very critical for proper treatment of the infected individuals as the sustained virological response of the standard antiviral interferon therapy is genotype dependent. We observed at our diagnostic center that 15.6% of HCV patient's samples were not genotype-able by using Ohno et al method. The genotyped samples showed that 3a (68.3%) is the major prevalent genotype in Pakistan followed by 2a (10.3%), 3b (2.6%), 1b (1.5%), 2b (1.2%) and 1a (0.5%). Presence of large number of untypable HCV variants in the current study highlights an important issue of health care setup in Pakistan. Untypable HCV cases create difficulties in treatment of these patients. The problem of routine diagnostics setup of Pakistan should be addressed on priority basis to facilitate the medical professionals in patient's treatment and to help in achieving the maximum sustained virological response.