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Answer added in ELISA6 Where can I get the orignial article in which I can find the specific experimental procedures of ELISA?Dear Liu. usually, the in-house made ELISA are first optimized by using the standard requirements and concentrations for each of the reagent used. So... [more]
Publications (19) View all
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Article: Crimean-Congo Hemorrhagic Fever Asia-2 Genotype, Pakistan
Emerging Infectious Diseases 06/2013; Vol. 19,(No. 6,). · 6.79 Impact Factor -
Article: Genetic analysis and epidemiology of Crimean Congo hemorrhagic fever viruses in Baluchistan province of Pakistan.
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ABSTRACT: BACKGROUND: Pakistan is considered as an endemic country for Crimean-Congo Hemorrhagic fever with numerous outbreaks and sporadic cases reported during the past two decades. Majority of cases are reported from Baluchistan province with subsequent transmissions to non-endemic regions mainly through infected animals directly or via infested ticks. We hereby describe the molecular investigations of CCHF cases reported during 2008 in Quetta city of Baluchistan province. METHODS: Serum Samples from 44 patients, with clinical signs of hemorrhagic fever attending a tertiary care hospital in Quetta city, were collected and tested for CCHF virus antigen and genomic RNA, using capture IgM EIA kit and standard RT-PCR assay, respectively. The partial S-gene fragments were directly sequenced to get information related to the prevailing CCHFV genotypes and their molecular epidemiology in Pakistan. RESULTS: Out of the total forty four, sixteen (36%) samples were found positive for CCHF IgM. Similarly, viral RNA was detected in six (16%) samples. Phylogenetic analysis revealed that all study viruses belong to genotype Asia-1 with closest similarity (99-100%) to the previously reported strains from Pakistan, Afghanistan and Iran. CONCLUSION: We conclude that CCHF virus remains endemic within Baluchistan and its neighboring regions of Afghanistan warranting a need of incessant surveillance activities.BMC Infectious Diseases 05/2013; 13(1):201. · 3.12 Impact Factor -
Article: Serotype Diversity of Astroviruses in Rawalpindi, Pakistan during 2009–2010
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ABSTRACT: Astroviruses are globally known enteropathogens causing gastroenteritis and diarrhea, with eight well defined serotypes. Epidemiological studies have recognized serotype-1 as the most common subtype but no such data is available in Pakistan. During 2009–2010, we found astroviruses in 41 out of 535 (7%) samples collected from hospitalized children. Thirty one strains belonged to serotype-1 and clustered into two distinct lineages. Serotype-3, -4 and -6 were detected with 97–98% genetic homology to Indian and Chinese strains.PLoS ONE 04/2013; · 4.09 Impact Factor -
Article: Characterization of non-polio enterovirus isolates from acute flaccid paralysis children in Pakistan reflects a new genotype of EV-107.
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ABSTRACT: Enteroviruses comprise a group of genetically diverse RNA viruses that usually cause respiratory illnesses, myocarditis, enteritis, meningitis, encephalitis and poliomyelitis. In this study, 23 non-polio enteroviruses, isolated during September-October 2009 from stool specimens of children suffering from acute flaccid paralysis (AFP) were subjected to serotype determination by using RIVM antiserum pools. Twenty-two isolates were identified as members of human enterovirus B (HEV-B) species except one that remained untypeable. Phylogenetic analysis of the untypeable isolate (PAKNIH-2009/23) based on the sequence of 5'UTR, capsid and 2A regions confirmed that this isolate is closely related to TN94-0349 strain of EV-107 serotype with 70.4% nucleotide similarity in VP1/2A junction, 79.4% in the capsid coding region and 89.1% in 5'UTR region respectively. Our data strongly suggests that PAKNIH-2009/23 is a genetically distinct strain of EV-107 currently circulating in Pakistan. This study also highlights the probable role of non-polio viral etiologies associated with AFP and needs to plan new strategies especially during the post polio eradication era.Virus Research 10/2012; · 2.94 Impact Factor -
Article: Identification of human parechovirus genotype, HPeV-12, in a paralytic child with diarrhea.
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ABSTRACT: New genotypes of human parechoviruses have been readily identified after improvement of diverse diagnostic tools. We hereby report the detection of a new genotype, HPeV 12, from a child presented with diarrhea and paralysis. The genetic variability of human parechoviruses has recently expanded defining 16 genotypes however data available covers only 11 genotypes. The present study was designed to determine the genetic characterization of human parechovirus identified in a child with gastroenteritis and acute flaccid paralysis (AFP). Stool samples are referred to Virology Department, NIH-Pakistan for the routine detection of enteroviruses and polioviruses through cell culture and RT-PCR. Five of isolates showing cytopathic effect on L20B cell line but negative for poliovirus were further explored for human parechovirus using multiple cell lines and RT-PCR. Human Coxsackie A virus type 2, 3, 6 and 20 were found in four samples whereas the fifth sample contained human parechovirus genotype 12. Efficient growth of human parechovirus was found on L20B cells while Vero and LLC-MK2 cells showed no apparent cytopathic effect. This study describes the detection of a new human parechovirus genotype (HPeV-12) in a paralytic child with diarrhea. Human parechoviruses are now considered as potential pathogens that may cause a number of serious clinical complications especially in infants and young children. These findings emphasize to conduct large scale epidemiological surveys in the country to understand their association with clinical diseases especially gastroenteritis, respiratory and neurological disorders.Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 09/2012; 55(4):339-42. · 3.12 Impact Factor
