An Epidemiological Study of Dengue Outbreak in Pakistan

In 2016, Hazara Division reported his major outbreak of Dengue fever. In this context, current epidemiological and serological survey conducted to highlight the actual burden of Dengue fever in cities of Hazara Division. Blood samples were taken from the total of 1462 suspected people for detection of Dengue antibodies. Among these patients, 1359 (93%) were found to be positive for Dengue, including 965 (71%) males and 394 (29%) females. Distribution in keeping the presence of antibodies shows 897 (66%) IgM positive people. Second most frequently seen antibodies were both IgG and IgM in 435 (32%) people. Presence of IgG antibodies was detected in 27 (2%) individuals. 1142 (84%) of Dengue positive people were not found to be symptomatic while rest of 217 (16%) observed with various symptoms. In this outbreak peak incidence of Dengue fever was observed in Manshera city. Although minimum was seen in Abbottabad city. To conclude, this might be the largest outbreak in the history of Hazara Division and second in Khyber Pakhtunkhwa. We recommend that policymakers and the government of Khyber Pakhtunkhwa desperately need to make efforts to prevent this mounting ratio of Dengue fever and implement the vector management policies by environmental measures and promote awareness in this area

Evaluation of Currently Available Molecular Assays and Performance of Sampling Approaches for Detection of Sars-Cov-2 RNA

OBJECTIVES This study aims to identify the essential characteristics of diagnostic tests for SARS-CoV-2 and to discuss the limitations of currently available tests and their impact on the test selection process. METHODOLOGY The current study was conducted at Mardan Medical Complex (MMC). One hundred nasopharyngeal-positive samples were collected from February to March 2021. Oropharyngeal swab OPS, sputum, and blood samples were collected from the participants to detect SARS-CoV-2 RNA. RNA extraction of SARS-CoV-2 was done using a BigFish auto extractor. A Qiagen Thermal Cycler was used for genome amplification. Five different molecular assays, namely COVSIGN (N gene) Spain, BGI (ORF1ab gene) China, Maccura(ORF1ab, E and N gene) China, R-GENE (RdRp and N genes) France and Genuru (N gene, S gene and ORF ab/1) were used. RESULTS100 % positivity was recorded in the sputum of all individuals, followed by 91 % OPS and 21% blood. The highest positivity rate for different genes was observed. ROC (Receiver operating characteristic curve) was developed through SPPS version 26.00 to compare the sensitivity and specificity. CONCLUSION By comparing the results of different diagnostic kits, it was found that BGI and Maccura are the most sensitive and specific for diagnostic purposes against COVID-19

Surveillance of molecular markers of antimalarial drug resistance in Plasmodium falciparum and Plasmodium vivax in Federally Administered Tribal Area (FATA), Pakistan

This molecular epidemiological study was designed to determine the antimalarial drug resistance pattern, and the genetic diversity of malaria isolates collected from a war-altered Federally Administered Tribal Area (FATA), in Pakistan. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai and Kurram agencies of FATA region between May 2017 and May 2018, and they underwent DNA extraction and amplification. The investigation of gene polymorphisms in drug resistance genes (dhfr, dhps, crt, and mdr1) of Plasmodium falciparum and Plasmodium vivax was carried out by pyrosequencing and Sanger sequencing, respectively. Out of 679 PCR-confirmed malaria samples, 523 (77%) were P. vivax, 121 (18%) P. falciparum, and 35 (5%) had mixed-species infections. All P. falciparum isolates had pfdhfr double mutants (C59R+S108N), while pfdhfr/pfdhps triple mutants (C59R+S108N+A437G) were detected in 11.5% of the samples. About 97.4% of P. falciparum isolates contained pfcrt K76T mutation, while pfmdr1 N86Y and Y184F mutations were present in 18.2% and 10.2% of the samples. P. vivax pvdhfr S58R mutation was present in 24.9% of isolates and the S117N mutation in 36.2%, while no mutation in the pvdhps gene was found. Pvmdr1 F1076L mutation was found in nearly all samples, as it was observed in 98.9% of isolates. No significant anti-folate and chloroquine resistance was observed in P. vivax; however, mutations associated with antifolate-resistance were found, and the chloroquine-resistant gene has been observed in 100% of P. falciparum isolates. Chloroquine and sulphadoxine-pyrimethamine resistance were found to be high in P. falciparum and low in P. vivax. Chloroquine could still be used for P. vivax infection but need to be tested in vivo, whereas a replacement of the artemisinin combination therapy for P. falciparum appears to be justified

Malaria prevalence in Pakistan: A systematic review and meta-analysis (2006–2021)

Malaria is one of the major public health issues globally. Malaria infection spreads through mosquito bites from infected female Anopheles mosquitoes. This study aims to conduct a systematic review and meta-analysis on malaria prevalence in Pakistan from 2006 to 2021. We searched PubMed, Science Direct, EMBASE, EMCare, and Google Scholar to acquire data on the prevalence of malaria infections. We performed a meta-analysis with a random-effects model to obtain the pooled prevalence of malaria, Plasmodium vivax, and Plasmodium falciparum. Meta-analysis was computed using R 4.1.2 Version statistical software. I2 and time series analysis were performed to identify a possible source of heterogeneity across studies. A funnel plot and the Freeman-Tukey Double Arcsine Transformed Proportion were used to evaluate the presence of publication bias. Out of the 315 studies collected, only 45 full-text articles were screened and included in the final measurable meta-analysis. Pooled malaria prevalence in Pakistan was 23.3%, with Plasmodium vivax, Plasmodium falciparum, and mixed infection rates of 79.13%, 16.29%, and 3.98%, respectively. Similarly, the analysis revealed that the maximum malaria prevalence was 99.79% in Karachi and the minimum was 1.68% in the Larkana district. Amazingly, this systematic review and meta-analysis detected a wide variation in malaria prevalence in Pakistan. Pakistan's public health department and other competent authorities should pay close attention to the large decrease in mosquito populations to curb the infection rate

Prevalence of molecular markers of sulfadoxine–pyrimethamine and artemisinin resistance in Plasmodium falciparum from Pakistan

Abstract Background In Pakistan, artesunate (AS) in combination with sulfadoxine–pyrimethamine (SP) is the recommended treatment for uncomplicated Plasmodium falciparum malaria. Monitoring molecular markers of anti-malarial drug resistance is crucial for early detection and containment of parasite resistance to treatment. Currently, no data are available on molecular markers of artemisinin resistance (K13 mutations) in P. falciparum isolates from Pakistan. In this study, the prevalence of mutations associated with SP and artemisinin resistance was estimated in different regions of Pakistan. Methods A total of 845 blood samples that were positive for malaria parasites by microscopy or rapid diagnostic test were collected from January 2016 to February 2017 from 16 different sites in Pakistan. Of these samples, 300 were positive for P. falciparum by PCR. Polymorphisms in the P. falciparum dihydrofolate reductase (pfdhfr) and dihydropteroate synthase (pfdhps) genes were identified by pyrosequencing while polymorphisms in the propeller domain of the pfk13 gene were identified by Sanger sequencing. Results The prevalence of the PfDHFR 108N and 59R mutations was 100% and 98.8%, respectively, while the prevalence of PfDHFR 50R and 51I mutations was 8.6%. No mutation was observed at PfDHFR position 164. In PfDHPS, the prevalence of mutations at positions 436, 437, and 613 was 9.9%, 45.2%, and 0.4%, respectively. No mutations were found at PfDHPS positions 540 and 581. The prevalence of double PfDHFR mutants (59R + 108N) ranged from 93.8% to 100%, while the prevalence of parasites having the PfDHFR 59R + 108N mutations in addition to the PfDHPS 437G mutation ranged from 9.5% to 83.3% across different regions of Pakistan. Nine non-synonymous and four synonymous mutations were observed in the PfK13 propeller domain, none of which correspond to mutations validated to contribute to artemisinin resistance. Conclusion The absence of the highly resistant PfDHFR/PfDHPS quintuple mutant parasites and the lack of PfK13 mutations associated with artemisinin resistance is consistent with AS + SP being effective in Pakistan