title: Detection of multiple cocirculating wild poliovirus type 1 lineages through environmental surveillance: impact and progress during 2011-2013 in Pakistan.
abstract: BACKGROUND
  The environmental surveillance has proven to be a useful tool to identify poliovirus circulation in different countries and was started in Pakistan during July 2009 to support the acute flaccid paralysis (AFP) surveillance system.
  
  
  METHODS
  Sewage samples were collected from 27 environmental sampling (ENV) sites and processed for poliovirus isolation through 2-phase separation method. Poliovirus isolates were identified as Sabin-like or wild type through real-time polymerase chain reaction (PCR). Wild-type strains were subjected to VP1 gene sequencing and phylogenetic analysis performed using MEGA 5.0.
  
  
  RESULTS
  During 2011-2013, a total of 668 samples were collected from 4 provinces that resulted in 40% of samples positive for wild poliovirus type-1 (WPV-1). None of the samples were positive for WPV-3. The areas with high frequency of WPV-1 detection were Karachi-Gadap (69%), Peshawar (82%), and Rawalpindi (65%), whereas the samples from Quetta and Sukkur remained negative for WPV during 2013. Phylogenetic analysis revealed 3 major clusters with multiple poliovirus lineages circulating across different country areas as well as in bordering areas of Afghanistan.
  
  
  CONCLUSIONS
  Environmental surveillance in Pakistan has been proven to be a powerful tool to detect WPV circulation in the absence of poliomyelitis cases in many communities. Our findings emphasize the need to continue and expand such surveillance activities to other high-risk areas in the country.
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Muhammad Masroor
authors.initials: MM
authors.lastname: Alam
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Shahzad
authors.initials: S
authors.lastname: Shaukat
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Salmaan
authors.initials: S
authors.lastname: Sharif
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Mehar
authors.initials: M
authors.lastname: Angez
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Adnan
authors.initials: A
authors.lastname: Khurshid
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Farzana
authors.initials: F
authors.lastname: Malik
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Lubna
authors.initials: L
authors.lastname: Rehman
authors.affiliation: WHO Regional Reference Laboratory for Polio Eradication Initiative, Department of Virology, National Institute of Health, Chak Shahzad, Islamabad, Pakistan.
authors.forename: Syed Sohail Zahoor
authors.initials: SS
authors.lastname: Zaidi
mesh_terms: D036022:Capsid Proteins / Q000235:genetics; D016000:Cluster Analysis; D004784:Environmental Monitoring; D006801:Humans; D010154:Pakistan; D010802:Phylogeny; D017955:Poliovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012722:Sewage / Q000821:virology*
chemical_list: D036022:Capsid Proteins; D012722:Sewage; C442540:VP1 protein, Poliovirus
keywords: Pakistan; environmental surveillance; molecular epidemiology; poliovirus