Detection of peste des petits ruminants virus in formalin-fixed tissues

Abstract

Peste des petits ruminants virus that causes a highly infectious and often fatal disease of sheep and goats is confirmed by various diagnostic techniques among them being isolation of the virus from cell culture systems, viral ribonucleic acid (RNA) detection by molecular assays, and viral antigen detection by immunocapture enzyme-linked immunosorbent assay (IC ELISA), immunohistochemistry (IHC), and AGAR gel test. Whereas most of the confirmatory diagnostic procedures require pathological samples to be stored frozen to preserve integrity of the peste des petits ruminants (PPR) virus RNA, samples for IHC tests are preserved in 10% formalin. In this study, nine formalin-fixed pathological samples from three goats suspected of PPR were processed for extraction of PPR viral RNA and analyzed for detection with real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay. The results showed that five out of the nine tested samples returned positive for presences PPR viral genome. This study has established that field pathological samples of PPR-suspected cases, collected and stored in 10% formalin for up 2 years, could be used for PPR virus RNA extraction for disease virus confirmation.

Confirmation and Sequence analysis of N gene of PPRV in South Xinjiang, China

“…It was confirmed that the herd was infected with PPRV when the outbreak occurred. In the study, it should be noted that PPRV were detected from the formalin-fixed tissues samples which have several advantages: to prevent the spread of the virus, do not need cold chain transport and archived pathological tissues samples can be used for retrospective study, without affecting detection of PPRV (Kihu et al 2015). Phylogenetic analysis showed that all involved Chinese strains in the study belonged to lineage Ⅳ, but in different sub-branches with Tibetan strains of 2007(Bao, et al, 2014, Wu et al, 2015.…”

The extraction of Peste Des Petits Ruminants Virus RNA from paraffin-embedded tissues using a modified extraction method

“…To detect canine distemper virus RNA in FFPE tissues, Liang et al (2012) used in situ hybridization (ISH) whereas Seimon et al (2013) used RT-PCR and ISH. Furthermore, detection of PPRV specific RNA in FFPE tissues by real time RT-PCR has been reported (Kihu et al, 2015). Detection of PPRV RNA in FFPE samples allow to use FFPE tissues in the confirmation of PPR in laboratories where fresh tissues are not available for extraction.…”

Peste Des Petit Ruminants

“…Effective disinfectant agents include alcohol, ether, phenol, sodium hydroxide and common detergents. In areas where PPR is endemic, the commonly employed control mechanism is vaccination (Kihu et al, 2015). The virus can survive for long periods of time in chilled or frozen tissues.…”

“…New animals should be quarantined for three weeks before allowing them to mix with the flock. In a case of PPR outbreak, animals with signs of PPR should be isolated immediately and sheep and goats around the outbreak area should be vaccinated as soon as possible (Blood et al, 1983;Diallo et al, 1995;Spickler and Roth, 2006;Kihu et al, 2015). Vaccination is the most effective way to gain control epidemic PPR.…”

“…This fortified vaccine is resistant to temperature as high as 45 °C for 14 days with negligible loss in efficacy. The use of this vaccine to protect small ruminants will lead to effective control of PPR in developing countries (Kihu et al, 2015).…”