{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T15:09:36Z","timestamp":1788188976795,"version":"build-2803163510"},"update-to":[{"DOI":"10.1371\/journal.pntd.0009802","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,10,5]],"date-time":"2021-10-05T00:00:00Z","timestamp":1633392000000}}],"reference-count":56,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2021,9,23]],"date-time":"2021-09-23T00:00:00Z","timestamp":1632355200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Pakistan Health Research Council","award":["34\/MCS\/RDC\/FJMU\/2971"],"award-info":[{"award-number":["34\/MCS\/RDC\/FJMU\/2971"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:p>Dengue has become endemic in Pakistan with annual recurrence. A sudden increase in the dengue cases was reported from Rawalpindi in 2016, while an outbreak occurred for the first time in Peshawar in 2017. Therefore, a multi-center study was carried out to determine the circulating dengue virus (DENV) serotypes and Chikungunya virus (CHIKV) co-infection in Lahore, Rawalpindi, and Peshawar cities in 2016\u201318. A hospital-based cross-sectional study was carried out in Lahore and Rawalpindi in 2016\u201318, while a community-based study was carried out in Peshawar in 2017. The study participants were tested for dengue NS1 antigen using an immunochromatographic device while anti-dengue IgM\/IgG antibodies were detected by indirect ELISA. All NS1 positive samples were used for DENV serotyping using multiplex real-time PCR assay. Additionally, dengue samples were tested for CHIKV co-infection using IgM\/IgG ELISA. A total of 6291 samples were collected among which 8.11% were NS1 positive while 2.5% were PCR positive. DENV-2 was the most common serotype (75.5%) detected, followed by DENV-1 in 16.1%, DENV-3 in 3.9% and DENV-4 in 0.7% while DENV-1 and DENV-4 concurrent infections were detected in 3.9% samples. DENV-1 was the predominant serotype (62.5%) detected from Lahore and Rawalpindi, while DENV-2 was the only serotype detected from Peshawar. Comorbidities resulted in a significant increase (p-value&lt;0.001) in the duration of hospital stay of the patients. Type 2 diabetes mellitus substantially (p-value = 0.004) contributed to the severity of the disease. Among a total of 590 dengue positive samples, 11.8% were also positive for CHIKV co-infection. Co-circulation of multiple DENV serotypes and CHIKV infection in Pakistan is a worrisome situation demanding the urgent attention of the public health experts to strengthen vector surveillance.<\/jats:p>","DOI":"10.1371\/journal.pntd.0009802","type":"journal-article","created":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T14:33:27Z","timestamp":1632494007000},"page":"e0009802","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":18,"title":["Dengue and Chikungunya virus co-infection in major metropolitan cities of provinces of Punjab and Khyber Pakhtunkhwa: A multi-center study"],"prefix":"10.1371","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8748-7741","authenticated-orcid":true,"given":"Faiz Ahmed","family":"Raza","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hasnain","family":"Javed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1408-069X","authenticated-orcid":true,"given":"Muhammad Mujeeb","family":"Khan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Obaid","family":"Ullah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Areeba","family":"Fatima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad","family":"Zaheer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6085-2390","authenticated-orcid":true,"given":"Saima","family":"Mohsin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shahida","family":"Hasnain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5658-3506","authenticated-orcid":true,"given":"Ruqyya","family":"Khalid","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arslan Ahmed","family":"Salam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,9,23]]},"reference":[{"key":"pntd.0009802.ref001","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/S0966-842X(01)02288-0","article-title":"Epidemic dengue\/dengue hemorrhagic fever as a public health, social and economic problem in the 21st century","volume":"10","author":"DJ Gubler","year":"2002","journal-title":"Trends in Microbiology"},{"key":"pntd.0009802.ref002","first-page":"457","volume-title":"Dengue vaccine: WHO position paper\u2013September","author":"WHO","year":"2018"},{"key":"pntd.0009802.ref003","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1126\/science.342.6157.415","article-title":"Surprising new dengue virus throws a spanner in disease control efforts","volume":"342","author":"D Normile","year":"2013","journal-title":"Science"},{"key":"pntd.0009802.ref004","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1126\/science.aan6836","article-title":"Antibody-dependent enhancement of severe dengue disease in humans","volume":"358","author":"LC Katzelnick","year":"2017","journal-title":"Science"},{"key":"pntd.0009802.ref005","volume-title":"Dengue and severe dengue","author":"WHO","year":"2018"},{"key":"pntd.0009802.ref006","first-page":"25","article-title":"Epidemiology of dengue in Pakistan, present prevalence and guidelines for future control","volume":"4","author":"S Ahmad","year":"2017","journal-title":"International Journal of Mosquito Research"},{"key":"pntd.0009802.ref007","volume-title":"The fauna of British India including Ceylon and Burma","author":"PJ Barraud","year":"1934"},{"key":"pntd.0009802.ref008","first-page":"16","article-title":"Arboviruses of public health importance in Pakistan","volume":"31","author":"CC Hayes","year":"1981","journal-title":"The Journal of the Pakistan Medical Association"},{"key":"pntd.0009802.ref009","first-page":"183","article-title":"The bionomics of the mosquitoes of the Changa Manga National forest, West Pakistan","volume":"1","author":"M Aslamkhan","year":"1969","journal-title":"Pakistan Journal of Zoology"},{"key":"pntd.0009802.ref010","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/0035-9203(82)90130-4","article-title":"West Nile virus in Pakistan. 1. Sero-epidemiological studies in Punjab Province","volume":"76","author":"CG Hayes","year":"1982","journal-title":"Transactions of the Royal Society of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref011","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1002\/jmv.1890260304","article-title":"Seroepidemiological study of infection with West Nile virus in Karachi, Pakistan, in 1983 and 1985","volume":"26","author":"M Sugamata","year":"1988","journal-title":"Journal of Medical Virology"},{"key":"pntd.0009802.ref012","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1016\/0035-9203(95)90412-3","article-title":"Dengue haemorrhagic fever outbreak in Karachi, Pakistan, 1994","volume":"89","author":"YC Chan","year":"1995","journal-title":"Transactions of the Royal Society of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref013","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/S1201-9712(98)90052-2","article-title":"Expansion of epidemic dengue viral infections to Pakistan","volume":"2","author":"RE Paul","year":"1998","journal-title":"International Journal of Infectious Diseases"},{"key":"pntd.0009802.ref014","doi-asserted-by":"crossref","first-page":"182","DOI":"10.3201\/eid1301.060376","article-title":"Dengue virus serotype 3, Karachi, Pakistan","volume":"13","author":"B Jamil","year":"2007","journal-title":"Emerging Infectious Diseases"},{"key":"pntd.0009802.ref015","doi-asserted-by":"crossref","first-page":"1114","DOI":"10.1016\/j.trstmh.2007.06.016","article-title":"Dengue outbreak in Karachi, Pakistan, 2006: experience at a tertiary care center","volume":"101","author":"E Khan","year":"2007","journal-title":"Transactions of the Royal Society of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref016","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.jcv.2008.06.003","article-title":"Co-circulations of two genotypes of dengue virus in 2006 out-break of dengue hemorrhagic fever in Karachi, Pakistan","volume":"43","author":"E Khan","year":"2008","journal-title":"Journal of Clinical Virology"},{"key":"pntd.0009802.ref017","doi-asserted-by":"crossref","first-page":"e54","DOI":"10.1016\/j.ijid.2009.10.008","article-title":"Multiple dengue serotypes and high frequency of dengue hemorrhagic fever at two tertiary care hospitals in Lahore during the 2008 dengue virus outbreak in Punjab, Pakistan","volume":"14","author":"MA Humayoun","year":"2010","journal-title":"International Journal of Infectious Diseases"},{"key":"pntd.0009802.ref018","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1186\/1471-2180-11-200","article-title":"Serotype and genotype analysis of dengue virus by sequencing followed by phylogenetic analysis using samples from three mini outbreaks-2007-2009 in Pakistan","volume":"11","author":"Z Fatima","year":"2011","journal-title":"BMC Microbiology"},{"key":"pntd.0009802.ref019","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1097\/MAJ.0b013e3182217001","article-title":"Prevalence and molecular characterization of dengue viruses serotypes in 2010 epidemic","volume":"343","author":"N Mahmood","year":"2012","journal-title":"The American Journal of the Medical Sciences"},{"key":"pntd.0009802.ref020","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1186\/s12985-016-0603-6","article-title":"Circulating serotypes of dengue virus and their incursion into non-endemic areas of Pakistan; a serious threat","volume":"13","author":"A Ali","year":"2016","journal-title":"Virology Journal"},{"key":"pntd.0009802.ref021","doi-asserted-by":"crossref","first-page":"151","DOI":"10.3343\/alm.2017.37.2.151","article-title":"Dengue virus serotypes circulating in Khyber Pakhtunkhwa Province, Pakistan, 2013\u20132015","volume":"37","author":"M Suleman","year":"2017","journal-title":"Annals of Laboratory Medicine"},{"key":"pntd.0009802.ref022","volume-title":"Provisional summary results of 6th population and housing census-2017","author":"Pakistan Bureau of Statistics","year":"2017"},{"key":"pntd.0009802.ref023","first-page":"1","volume-title":"Revised Criteria for Diagnosis of Dengue Fever (Dengue fever during Rainy Season)","author":"Dengue Expert Advisory Group","year":"2014"},{"key":"pntd.0009802.ref024","volume-title":"Revised and expanded edition","author":"WHO","year":"2011"},{"key":"pntd.0009802.ref025","first-page":"1","volume-title":"Deaths from dengue in Pakistan","author":"WHO","year":"2017"},{"key":"pntd.0009802.ref026","doi-asserted-by":"crossref","first-page":"3781","DOI":"10.1128\/JCM.01870-14","article-title":"Comparison of two commercial real-time PCR assays for detection of dengue virus in patient serum samples","volume":"52","author":"J Saengsawang","year":"2014","journal-title":"Journal of Clinical Microbiology"},{"key":"pntd.0009802.ref027","volume-title":"The Rutherford","author":"AITbiotech","year":"2020"},{"key":"pntd.0009802.ref028","doi-asserted-by":"crossref","first-page":"e0004119","DOI":"10.1371\/journal.pntd.0004119","article-title":"Utilization of an eilat virus-based chimera for serological detection of chikungunya infection","volume":"9","author":"JH Erasmus","year":"2015","journal-title":"PLOS Neglected Tropical Diseases"},{"key":"pntd.0009802.ref029","doi-asserted-by":"crossref","first-page":"182","DOI":"10.4269\/ajtmh.16-0013","article-title":"Evaluation of commercially available Chikungunya virus immunoglobulin M Detection assays","volume":"95","author":"BW Johnson","year":"2016","journal-title":"The American Journal of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref030","doi-asserted-by":"crossref","first-page":"16055","DOI":"10.1038\/nrdp.2016.55","article-title":"Dengue infection","volume":"2","author":"MG Guzman","year":"2016","journal-title":"Nature Reviews Disease Primers"},{"key":"pntd.0009802.ref031","doi-asserted-by":"crossref","first-page":"495","DOI":"10.4269\/ajtmh.1974.23.495","article-title":"Fatal hemorrhagic disease and shock associated with primary dengue infection on a Pacific island","volume":"23","author":"WJ Barnes","year":"1974","journal-title":"The American Journal of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref032","doi-asserted-by":"crossref","first-page":"e0004398","DOI":"10.1371\/journal.pntd.0004398","article-title":"Incomplete Protection against Dengue Virus Type 2 Re-infection in Peru","volume":"10","author":"BM Forshey","year":"2016","journal-title":"PLOS Neglected Tropical Diseases"},{"key":"pntd.0009802.ref033","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1093\/infdis\/jiw099","article-title":"Homotypic dengue virus reinfections in Nicaraguan children","volume":"214","author":"JJ Waggoner","year":"2016","journal-title":"The Journal of Infectious Diseases"},{"issue":"Suppl 1","key":"pntd.0009802.ref034","doi-asserted-by":"crossref","first-page":"E48","DOI":"10.1111\/j.1749-6632.2009.05052.x","article-title":"Antibodies determine virulence in dengue","volume":"1171","author":"SB Halstead","year":"2009","journal-title":"Annals of the New York Academy of Sciences"},{"key":"pntd.0009802.ref035","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1093\/infdis\/jit436","article-title":"A shorter time interval between first and second dengue infections is associated with protection from clinical illness in a school-based cohort in Thailand","volume":"209","author":"KB Anderson","year":"2014","journal-title":"The Journal of Infectious Disease"},{"key":"pntd.0009802.ref036","doi-asserted-by":"crossref","first-page":"11887","DOI":"10.1073\/pnas.1504964112","article-title":"Impact of human mobility on the emergence of dengue epidemics in Pakistan","volume":"112","author":"A Wesolowski","year":"2015","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"pntd.0009802.ref037","doi-asserted-by":"crossref","first-page":"307","DOI":"10.5099\/aj120400307","article-title":"Dengue virus serotype 2 (DEN-2): the causative agent of 2011-dengue epidemic in Pakistan","volume":"4","author":"M Idrees","year":"2012","journal-title":"American Journal of Biomedical Sciences"},{"key":"pntd.0009802.ref038","doi-asserted-by":"crossref","first-page":"245","DOI":"10.2217\/fvl-2017-0155","article-title":"Analysis of dengue virus burden and serotypes pattern in Faisalabad, 2016\u20132017","volume":"13","author":"M Yousaf","year":"2018","journal-title":"Future Virology"},{"key":"pntd.0009802.ref039","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.apjtm.2016.01.013","article-title":"Concurrent infections of dengue viruses serotype 2 and 3 in patient with severe dengue from Jakarta, Indonesia","volume":"9","author":"S Lardo","year":"2016","journal-title":"Asian Pacific Journal of Tropical Medicine"},{"key":"pntd.0009802.ref040","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1186\/s13104-019-4164-9","article-title":"Concurrent infections of dengue virus serotypes in Bali, Indonesia","volume":"12","author":"S Masyeni","year":"2019","journal-title":"BMC Research Notes"},{"key":"pntd.0009802.ref041","doi-asserted-by":"crossref","first-page":"e0074","DOI":"10.1590\/0074-02760180082","article-title":"Comorbidities increase in-hospital mortality in dengue patients in Brazil","volume":"113","author":"GL Werneck","year":"2018","journal-title":"Memorias do Instituto Oswaldo Cruz"},{"key":"pntd.0009802.ref042","doi-asserted-by":"crossref","first-page":"e0004619","DOI":"10.1371\/journal.pntd.0004619","article-title":"NTD and NCD co-morbidities: the example of dengue fever","volume":"10","author":"P Mehta","year":"2016","journal-title":"Plos Neglected Tropical Diseases"},{"key":"pntd.0009802.ref043","doi-asserted-by":"crossref","first-page":"e2379","DOI":"10.1089\/vbz.2018.2379","article-title":"Admission clinicopathological factors associated with prolonged hospital stay among hospitalized patients with dengue viral infections","volume":"19","author":"SS Willeam Peter","year":"2019","journal-title":"Vector Borne and Zoonotic Diseases"},{"key":"pntd.0009802.ref044","doi-asserted-by":"crossref","first-page":"e1641","DOI":"10.1371\/journal.pntd.0001641","article-title":"Diabetes with hypertension as risk factors for adult dengue hemorrhagic fever in a predominantly dengue serotype 2 epidemic: a case control study","volume":"6","author":"J Pang","year":"2012","journal-title":"PloS Neglected Tropical Diseases"},{"key":"pntd.0009802.ref045","doi-asserted-by":"crossref","first-page":"e0081060","DOI":"10.1371\/journal.pone.0081060","article-title":"Risk factors for fatality among confirmed adult dengue inpatients in Singapore: a matched case-control study","volume":"8","author":"TL Thein","year":"2013","journal-title":"PLoS One"},{"key":"pntd.0009802.ref046","doi-asserted-by":"crossref","first-page":"e0200200","DOI":"10.1371\/journal.pone.0200200","article-title":"Prevalence of chronic comorbidities in dengue fever and West Nile virus: A systematic review and meta-analysis","volume":"13","author":"A Badawi","year":"2018","journal-title":"PLoS One"},{"key":"pntd.0009802.ref047","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1016\/S1473-3099(16)00026-8","article-title":"The global burden of dengue: an analysis from the Global Burden of Disease Study 2013","volume":"16","author":"JD Stanaway","year":"2016","journal-title":"The Lancet Infectious Diseases"},{"key":"pntd.0009802.ref048","doi-asserted-by":"crossref","first-page":"670","DOI":"10.4269\/ajtmh.11-0774","article-title":"Determinants of mortality from severe dengue in Brazil: a population-based case-control study","volume":"88","author":"GH Moraes","year":"2013","journal-title":"The American Journal of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref049","doi-asserted-by":"crossref","first-page":"1022","DOI":"10.1111\/j.1365-3156.2004.01295.x","article-title":"Risk factors and clinical features associated with severe dengue infection in adults and children during the 2001 epidemic in Chonburi, Thailand","volume":"9","author":"O Wichmann","year":"2004","journal-title":"Tropical Medicine and International Health"},{"key":"pntd.0009802.ref050","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.virol.2009.07.024","article-title":"Replication cycle of chikungunya: a re-emerging arbovirus","volume":"393","author":"M Solignat","year":"2009","journal-title":"Virology"},{"key":"pntd.0009802.ref051","volume-title":"Chikungunya virus","author":"CDC","year":"2019"},{"key":"pntd.0009802.ref052","doi-asserted-by":"crossref","first-page":"1708","DOI":"10.3390\/ijms18081708","article-title":"Co-Infection of mosquitoes with Chikungunya and Dengue viruses reveals modulation of the replication of both viruses in midguts and salivary glands of Aedes aegypti mosquitoes","volume":"18","author":"A Le Coupanec","year":"2017","journal-title":"International Journal of Molecular Sciences"},{"key":"pntd.0009802.ref053","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/cid\/cis530","article-title":"Recent introduction and rapid dissemination of Chikungunya virus and Dengue virus serotype 2 associated with human and mosquito coinfections in Gabon, central Africa","volume":"55","author":"M Caron","year":"2012","journal-title":"Clinical Infectious Diseases"},{"key":"pntd.0009802.ref054","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1016\/0035-9203(83)90106-2","article-title":"A sero-epidemiological survey for certain arboviruses (Togaviridae) in Pakistan","volume":"77","author":"MA Darwish","year":"1983","journal-title":"Transactions of the Royal Society of Tropical Medicine and Hygiene"},{"key":"pntd.0009802.ref055","volume-title":"Mysterious disease affects 30,000 people in Karachi","author":"D News","year":"2016"},{"key":"pntd.0009802.ref056","doi-asserted-by":"crossref","first-page":"e0007118","DOI":"10.1371\/journal.pntd.0007118","article-title":"Epidemiological trend of chikungunya outbreak in Pakistan: 2016\u20132018","volume":"13","author":"N Badar","year":"2019","journal-title":"PLOS Neglected Tropical Diseases"}],"updated-by":[{"DOI":"10.1371\/journal.pntd.0009802","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,10,5]],"date-time":"2021-10-05T00:00:00Z","timestamp":1633392000000}}],"container-title":["PLOS Neglected Tropical Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0009802","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T14:34:32Z","timestamp":1632494072000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0009802"}},"subtitle":[],"editor":[{"given":"Olaf","family":"Horstick","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2021,9,23]]},"references-count":56,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9,23]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pntd.0009802","relation":{"new_version":[{"id-type":"doi","id":"10.1371\/journal.pntd.0009802","asserted-by":"object"}]},"ISSN":["1935-2735"],"issn-type":[{"value":"1935-2735","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,23]]}}}