Preparation of a house ELISA kit for detecting Peste des petits ruminants Virus (PPRV) antibodies

Hosny, Wafaa Abd El Wahab; Baheeg, Eman; Hassanein, Sozan; Mohamed, S. A.

doi:10.21608/bvmj.2021.84188.1446

Cited by 2 publications

(4 citation statements)

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“…Indirect IELISA, which performed much better than cELISA, presented 150 positives out of 176 serum samples (Table 4). The 90.60% sensitivity and 85.23% specificity (Table 4) rates are consistent with the results presented by Hosny and colleagues in their study [8]. With IELISA, a large number of serum samples can be rapidly and affordably tested.…”

Section: Discussionsupporting

confidence: 90%

“…Maximum three consecutive blind passages on Vero cells were required to successfully isolate PPRV from field tissue samples; the virus was then identified by its characteristic cytopathic effects (CPE) (O.I.E., 2013) [33]. According to Hosny and colleagues (2021) [8], the PPRV virus is presently spreading to new hosts. Recently, camel deaths occurred in Sudan from a fatal respiratory disease that was traced back to PPRV; however, sheep and goats are the most common hosts for PPRV.…”

Section: Discussionmentioning

confidence: 99%

“…The suspected outbreaks of PPR reported by resource persons within three different regions including Islamabad territory (33.6844° N, 73.0479°), Fateh Jang (33.8611° N, 72.4140° E), and Gilgit (35.9208° N, 74.3082° E) were attended,during 2020-2021.The affected herds were visited to record the clinical manifestations and clinical specimen were collected following standard procedure [8,9]. The sampled animals had no history of vaccination against PPRV.The sampling locations were presented in (Figure 1).…”

Section: Clinical Specimen Collectionmentioning

confidence: 99%

“…Camels are not the only animals that have been affected by the disease [7]. According to Hosny and colleagues [8] and others [9], who investigated the recent deaths of camels in Sudan from a fatal respiratory disease that was traced to PPRV, the PPR virus is spreading to new hosts, although sheep and goats are the most common hosts for PPRV.…”

Section: Introductionmentioning

confidence: 99%

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The Molecular Characterization of Lineage-IV Peste Des Petits Ruminants Virus and the Development of In-House IELISA for Its Rapid Detection

Rana,

Khan,

Hassan

et al. 2023

Preprint

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Peste des Petits Ruminants (PPR), a highly contagious viral disease, causes significant economic losses in sheep and goats. Laboratory diagnosis is crucial to disease control and eradication. Since PPRVs have recently adopted new hosts, genetic diversity within isolates of the same lineage is more likely than if they were host-specific, sheep and goats most often. ELISA detects antibodies and antigens in serology. However, mishandling, environmental conditions (temperature and humidity), storage, and sensitivity issues of commercially available ELISA kits prevent rapid PPR virus detection in third-world countries like Pakistan. In the 2020-21 outbreaks in Pakistan, 325 blood samples, 19 swabs, and 6 virus tissue samples of sheep and goats were collected from Gilgit, Islamabad, and Fateh Jang. These virus isolates were submitted to NCBI for partial N gene sequencing. Antigen was prepared from indigenous virus Using semi-purified antigen from PPRV, an indirect ELISA for the detection of PPR antibodies in goat and sheep serum was developed using MW600922 grown in Vero cells. Dilutions of 1:200 serums and 1:32 antigens improve antibody detection. The N-gene-based analysis revealed a better understanding of PPRV genetic characterization. This study develops an RT-PCR assay to detect PPRV by targeting the N-protein gene. This assay offers an accurate and affordable diagnostic tool for PPRV and partial genome sequencing. Results described that PPRV isolates from the current study have 99.73% and 99.4% similarity to previously published Lahore and Faisalabad isolates from Pakistan, respectively. Compared to previous isolates from NCBI data, the virus showed high divergence rates with the Turkish strain. The comparative analysis between Commercial kit (c-ELISA) and I-ELISA revealed that the former had a high sensitivity of 90.60% and specificity of 85.23% compared with the cELISA kit. By comparing I-ELISA and VNT, we find that the current assay is 100% specific and 82.14 % sensitive. Based on these results, serological surveys for PPR antibodies, on a larger scale, in small ruminants can be conducted with indirect ELISA rather than competitive ELISA. Furthermore, the cost and storage efficiency was highly significant because the currently developed method has a low production cost, which is much lesser than the commercially available kit. Our findings demonstrated a significant breakthrough in terms of cost-effectiveness and storage efficiency, and they are highly recommended for developing countries.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Section: Clinical Specimen Collectionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Molecular Characterization of Lineage-IV Peste Des Petits Ruminants Virus and the Development of In-House IELISA for Its Rapid Detection

Rana,

Khan,

Hassan

et al. 2023

Preprint

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Molecular Characterization of Lineage-IV Peste Des Petits Ruminants Virus and the Development of In-House Indirect Enzyme-Linked Immunosorbent Assay (IELISA) for its Rapid Detection”

Kamal,

Khan,

Hassan

et al. 2024

Biol Proced Online

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Peste des petits ruminants (PPRV), a highly contagious viral disease, causes significant economic losses concerning sheep and goats. Recently, PPR viruses (PPRVs), have adopted new hosts and lineage IV of PPRVs represents genetic diversity within the same lineage. 350 samples, including blood, swabs, and tissues from sheep/goats, were collected during the 2020–2021 disease outbreaks in Pakistan. These samples were analysed through RT-PCR and three isolates of PPRV with accession numbers, MW600920, MW600921, and MW600922, were submitted to GenBank, based on the partial N-gene sequencing. This analysis provides a better understanding of genetic characterizations and a targeted RT-PCR approach for rapid PPRV diagnosis. An IELISA test was developed using the semi-purified antigen MW600922 isolate grown in Vero cells. The PPRV isolates currently present high divergence with the Turkish strain; conversely, similarities equivalent to 99.73% were observed for isolates collected from Pakistan. The developed indirect ELISA (IELISA) test demonstrated antibody detection rates at dilutions of 1:200 for antibodies (serum) and 1:32 for antigens. In comparison to cELISA, high specificity (85.23%) and sensitivity (90.60%) rates were observed. In contrast to the virus neutralization test (VNT), IELISA was observed to be 100% specific and 82.14% sensitive in its results. Based on these results, serological surveys conducted for PPR antibodies using IELISA can be a more effective strategy on a larger scale. Furthermore, our results demonstrate a significant breakthrough in the research in terms of cost-effectiveness and storage efficiency, and the developed IELISA test is highly recommended for use in developing countries.

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Preparation of a house ELISA kit for detecting Peste des petits ruminants Virus (PPRV) antibodies

Cited by 2 publications

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The Molecular Characterization of Lineage-IV Peste Des Petits Ruminants Virus and the Development of In-House IELISA for Its Rapid Detection

The Molecular Characterization of Lineage-IV Peste Des Petits Ruminants Virus and the Development of In-House IELISA for Its Rapid Detection

Molecular Characterization of Lineage-IV Peste Des Petits Ruminants Virus and the Development of In-House Indirect Enzyme-Linked Immunosorbent Assay (IELISA) for its Rapid Detection”

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