Serodiagnosis Using Recombinant Nipah Virus Nucleocapsid Protein Expressed in
            <i>Escherichia coli</i>

Yu, Fangfang; Khairullah, Nor Shahidah; Inoue, Satoshi; Balasubramaniam, Vijayamalar; Berendam, Stella J.; Teh, Leok Kin; Ibrahim, N.; Rahman, Sohayati Abdul; Hassan, Sharifah Syed; Hasebe, Futoshi; Sinniah, M; Morita, Kouichi

doi:10.1128/jcm.00693-06

Cited by 51 publications

(33 citation statements)

References 24 publications

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“…However, its use is limited to BSL4 laboratories. To overcome this problem, recombinant proteins have been developed and used as an alternative antigen for serological detections of Henipaviruses [7,24,102]. A recombinant N protein ELISA was developed and used for screening the pig serum samples at HSADL.…”

Section: Diagnostic Tests and Facilitiesmentioning

confidence: 99%

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Nipah virus infection: current scenario

et al. 2013

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Section: Diagnostic Tests and Facilitiesmentioning

confidence: 99%

“…Institute of Tropical Medicine, Japan [102] 13. Neutralization assays for differential Henipavirus serology using Bio-Plex Protein Array Systems CSIRO, Australia [4] 14.…”

Section: Concluding Comments and Lessons To Be Learned From Nipah Expmentioning

confidence: 99%

Nipah virus infection: current scenario

et al. 2013

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“…Western blot analysis with MAb against the PPRVNP/ polyclonal antibodies or serum indicated that the bands observed in SDS-PAGE are virus specific. Such His-tag fusion protein has been expressed in case of PPR and Nipah virus also (Yu et al, 2006;Yadav et al, 2009). …”

Section: Discussionmentioning

confidence: 99%

“…This method has been successfully used for purification of several other expressed proteins (Yu et al, 2006;Sun et al, 2007;Yadav et al, 2009). Presence of N-terminal His-tag in the vector as well as C-terminal His-tag both in cloned gene product and expression vector facilitated easy and efficient Ni-NTA column protein purification.…”

Section: Discussionmentioning

confidence: 99%

“…Earlier recombinant antigen based diagnostic methods have been developed for the detection of several morbilliviruses such as rinderpest virus (RPV) (Kamata et al, 1993), PPRV (Ismail et al, 1995;Libeau et al, 1995;Choi et al, 2005b;Dechamma et al, 2006;Balamurugan et al, 2006;Yadav et al, 2009), canine distemper virus (CDV) (Latha et al, 2007) and Nipah virus (Yu et al, 2006). Among the structural proteins of morbilliviruses, N protein is the highly conserved immunogenic core protein (Lefebvre et al, 1991) and is expressed at a high level in infected cells (Diallo et al, 1994).…”

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Development of Recombinant Nucleocapsid Protein based indirect ELISA for Serodiagnosis of Peste des Petits Ruminants in Sheep and Goats

Balamurugan¹,

Roy²,

SowjanyaKumari³

et al. 2016

Adv. Anim. Vet. Sci.

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| In this study, expression of immunogenic portion of Peste des Petits ruminants virus (PPRV) nucleocapsid (N) protein in Escherichia coli (BL21) was envisaged to evaluate the potential use of recombinant protein as a diagnostic antigen in polyclonal antibodies based indirect ELISA for serodiagnosis. The immunogenic region of N gene coding sequences from PPR vaccine virus (Sungri 96 strain) was amplified, cloned in pET32a vector and expressed in E. coli as fusion protein for bulk production and easy Ni-NTA His-tag purification. The recombinant PPRV N protein (rPPRVNP) was expressed in E. coli at an optimal temperature of 37 °C with 1 mM IPTG for 5 h post induction and characterized by SDS-PAGE and western blot using PPRV-specific monoclonal and polyclonal antibodies or serum or anti-His-tag conjugate that confirmed PPRV specific protein with a size of ~50kDa. The expressed rPPRVNP was in insoluble form and was purified under denaturation condition by Ni-NTA purification method followed by refolding, renaturation methods and further concentrated by protein cut-off concentrators for obtaining single protein band. The rPPRVNP was assessed for its immunoreactivity as diagnostic antigen by immunoblotting and ELISA using standard PPRV specific antibodies. The immunogenic reactivity of expressed rPPRVNP was optimized in indirect ELISA using known true positive and negative sera with respect to PPRV antibodies. On standardization of rPPRVNP based indirect ELISA using 661serum samples, the relative diagnostic sensitivity and specificity of the assay 83.76% and 83.13% respectively was observed at cut off level of 25 Per cent Positivity (PP) with an agreement of Cohen's kappa value of 0.648 with good agreement. This indirect ELISA as additional diagnostic tool for diagnosis of PPR in sheep and goats and rPPRVNP could be a sustainable source of safe antigen in countries of non-endemicity without the need to handle infectious virus for sero-diagnosis.

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H enipaviruses

Aljofan

2015

Encyclopedia of Life Sciences

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Henipaviruses are the deadliest human pathogens of the Paramyxoviridae family. The genus Henipavirus includes two closely related, zoonotic, and highly pathogenic paramyxoviruses, Nipah virus (NiV) and Hendra virus (HeV), which cause high morbidity and mortality in animals and humans. There are no approved therapeutics to treat or prevent henipavirus infections in humans. The bat species Pteropus are the natural reservoir of henipaviruses, and zoonotic transmission can occur directly or through an intermediate host; horses for HeV and pigs for NiV. The unavailability of a treatment for these infections, the recent detection of HeV antibodies in family pets and the emergence of NiV infections in new geographic areas (Philippines) are an illustration of the risk that these viruses possess and their momentum of becoming a global threat. Unless an effective therapeutic against these viruses is developed, they will continue to emerge and cause significant challenges and threats in the coming years. Key Concepts The newly established genus Henipavirus of Paramyxoviridae family has three members Hendra, Nipah and Cedar viruses. Fruit bats of the genus Pteropus have been identified as the natural reservoir for henipaviruses. Both Nipah and Hendra viruses are highly pathogenic agents that cause acute encephalitis and respiratory illness with high mortality rate in humans, pigs and horses. There is no approved therapeutics for the treatment of henipaviruses, and they continue to pose a threat in Australia (Hendra virus) and East and Southeast Asia (Nipah virus) with a great potential to spread beyond these regions. M 102.4 is an experimental monoclonal antibody that targets the viral entry stage of Henipavirus and was proven to treat henipavirus infections in animals.

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Serodiagnosis Using Recombinant Nipah Virus Nucleocapsid Protein Expressed in Escherichia coli

Cited by 51 publications

References 24 publications

Nipah virus infection: current scenario

Nipah virus infection: current scenario

Development of Recombinant Nucleocapsid Protein based indirect ELISA for Serodiagnosis of Peste des Petits Ruminants in Sheep and Goats

H enipaviruses

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