A robust lentiviral pseudotype neutralisation assay for in-field serosurveillance of rabies and lyssaviruses in Africa

Abstract

The inflexibility of existing serological techniques for detection of rabies in surveillance constrains the benefit to be gained from many current control strategies. We analysed 304 serum samples from Tanzanian dogs for the detection of rabies antibodies in a pseudotype assay using lentiviral vectors bearing the CVS-11 envelope glycoprotein. Compared with the widely used gold standard fluorescent antibody virus neutralisation assay, a specificity of 100% and sensitivity of 94.4% with a strong correlation of antibody titres (r = 0.915) were observed with the pseudotype assay. To increase the assay's surveillance specificity in Africa we incorporated the envelope glycoprotein of local viruses, Lagos bat virus, Duvenhage virus or Mokola virus and also cloned the lacZ gene to provide a reporter element. Neutralisation assays using pseudotypes bearing these glycoproteins reveal that they provide a greater sensitivity compared to similar live virus assays and will therefore allow a more accurate determination of the distribution of these highly pathogenic infections and the threat they pose to human health. Importantly, the CVS-11 pseudotypes were highly stable during freeze–thaw cycles and storage at room temperature. These results suggest the proposed pseudotype assay is a suitable option for undertaking lyssavirus serosurveillance in areas most affected by these infections.

The use of equine influenza pseudotypes for serological screening

“…These include Ebola virus (Wool-Lewis and Bates, 1998; Ito et al, 2001), hepatitis C (Bartosch et al, 2003; Hsu et al, 2003), SARS coronavirus (Nie et al, 2004; Temperton et al, 2005), rabies and lyssaviruses (Wright et al, 2008; 2009; 2010) as well as avian influenza viruses (Temperton et al, 2007). In contrast to the native viruses, their pseudotype counterparts can be handled in BSL1/2 facilities.…”

Lentiviral hepatitis B pseudotype entry requires sodium taurocholate co-transporting polypeptide and additional hepatocyte-specific factors

“…This system has been used to study the entry pathways of a range of viruses, including human immunodeficiency virus (HIV), hepatitis C virus (HCV), pathogenic avian influenza and Ebola, offering a rapid and reproducible analysis of the viral glycoprotein-receptor interactions (Moller-Tank & Maury, 2015;Temperton et al, 2007;Wright et al, 2009;Zimmer et al, 2014). Previous reports showed that lentiviral HBVpps were infectious for human hepatocytes; however, given the limited knowledge of internalization pathways at the time, the receptor dependency was not studied (Chai et al, 2007;Sung & Lai, 2002).…”

“…The HBV surface glycoprotein is unlike other viral glycoproteins tested to date (Moller-Tank & Maury, 2015;Temperton et al, 2007;Wright et al, 2009;Zimmer et al, 2014) in its failure to produce infectious pseudoparticles in human embryonal kidney cells (293T), requiring a liver cell background for infectious particles genesis. Our results show that whilst NTCP does play an essential role in HBVpp entry, it is unlikely to be the sole receptor facilitating infection.…”

Lyophilisation of influenza, rabies and Marburg lentiviral pseudotype viruses for the development and distribution of a neutralisation -assay-based diagnostic kit

“…"Advantageous"qualities"of" 243" the"PVNA"platform"include"the"ability"to"conduct"the"assay"in"BSLS1"laboratories,"as"well"as"the" 244" availability"to"perform"multiplex"assays"and"incorporate"'lowScost'"reporter"genes"into" 245" pseudoparticles,"which"all"reduce"the"cost"implications"and"increase"the"potential"ubiquity"of" 246" pseudotyping" (Wright"et"al.,"2009(Wright"et"al.," ,"2010"Kaku"et"al.,"2012). ""However,"the"current"necessity"to"store" 247" aliquots"of"PV"supernatant"at"S80°C"and"to"maintain"the"coldSchain"during"PV"transportation"present" 248" serious"monetary"obstacles"for"laboratories"to"acquire"such"reagents,"especially"if"on"a"limited" 249" budget.""…”