Recombinase-Aided Amplification Coupled with Lateral Flow Dipstick for Efficient and Accurate Detection of Porcine Parvovirus

He, Ya‐Ling; Chen, Wenxian; Fan, Jindai; Fan, Songqing; Ding, Hongxing; Chen, Jinding; Lin, Yan

doi:10.3390/life11080762

Cited by 9 publications

(10 citation statements)

References 25 publications

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“…Four BoV sequences (GenBank Accession No. OR682586-OR682589) detected in the duodenum of two animals (656DU-2022 and 657DU-2022) and in the liver and intestinal samples of an additional hedgehog (655DU-2022 and 655L-2022) showed the highest nt (98.9–99.4%) and aa (100%) identities to porcine bocavirus (PBoV) strains detected in pigs in China [ 23 , 24 , 25 ], South Korea [ 26 ], Belgium [ 27 ], East Africa [ 28 ], and the USA [ 29 ]. Furthermore, a high nt identity, ranging from 87.3% to 90.4%, was found for sequences related to PBoVs but detected in rats, minks, and mice [ 30 , 31 , 32 , 33 , 34 ].…”

Section: Resultsmentioning

confidence: 99%

Molecular Surveillance for Bocaparvoviruses and Bufaviruses in the European Hedgehog (Erinaceus europaeus)

Sarchese,

Palombieri,

Prandi

et al. 2024

Microorganisms

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The presence of bocaparvoviruses (BoVs) and bufaviruses (BuVs) in the European hedgehog (Erinaceus europaeus) was investigated by screening duodenal and liver samples collected from 183 carcasses, delivered to wildlife rescue centers located in northwestern Italy. BoV DNA was detected in 15 animals (8.2%), with prevalences of 7.1% (13/183) and 2.7% (5/183) in intestine and liver samples, respectively. Upon the sequence analyses of the NS1 gene, two highly divergent BoVs (65.5–67.8% nt identities) were identified. Fourteen strains showed the highest identity (98.3–99.4% nt) to the hedgehog BoV strains recently detected in China in Amur hedgehogs (Erinaceus amurensis), whilst four strains were genetically related (98.9–99.4% nt identities) to the porcine BoVs identified in pigs and classified in the species Bocaparvovirus ungulate 4, which included related viruses also found in rats, minks, shrews, and mice. BuV DNA was detected in the duodenal samples of two hedgehogs, with a prevalence rate of 1.1%. The nearly full-length genome of two BuV strains, Hedgehog/331DU-2022/ITA and Hedgehog/1278DU/2019/ITA, was reconstructed. Upon phylogenetic analysis based on the NS and VP aa sequences, the Italian hedgehog BuVs tightly clustered with the BuVs recently identified in the Chinese Amur hedgehogs, within a potential novel candidate species of the genus Protoparvovirus.

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Section: Resultsmentioning

confidence: 99%

Molecular Surveillance for Bocaparvoviruses and Bufaviruses in the European Hedgehog (Erinaceus europaeus)

Sarchese,

Palombieri,

Prandi

et al. 2024

Microorganisms

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“…To identify the causative agent of diarrheic piglets, faecal samples were examined by RT‐PCR for common porcine enteric viruses, including transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhoea virus (PEDV), porcine deltacoronavirus (PDCoV), genogroup A and C rotavirus (RVA and RVC), porcine astrovirus (PAstV), porcine sapovirus (PSaV), porcine norovirus (PNoV), mammalian orthoreovirus (MRV) and porcine parvovirus (PPV), using specific primers (Table S1) as previously described (Ding et al., 2020; He et al., 2021; Kim et al., 2006; Kim et al., 2010; Lelli et al., 2013; Reuter et al., 2011; Wang et al., 2006). Faecal sample tested negative for above enteric viruses was subjected to virus isolation.…”

Section: Methodsmentioning

confidence: 99%

“…and porcine parvovirus (PPV), using specific primers (Table S1) as previously described (Ding et al, 2020;He et al, 2021;Kim et al, 2006;Kim et al, 2010;Lelli et al, 2013;Reuter et al, 2011;Wang et al, 2006). Faecal sample tested negative for above enteric viruses was subjected to virus isolation.…”

Section: Isolation Of Piv5 Strainmentioning

confidence: 99%

Characterization of parainfluenza virus 5 from diarrheic piglet highlights its zoonotic potential

Ibrahim

Zhang

Werid

et al. 2022

Transbounding Emerging Dis

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Parainfluenza virus 5 (PIV5), a member of paramyxoviruses, causes respiratory and neurological infection in several animal species. Whereas information on PIV5 infection in digestive system is very scarce. Here, we successfully isolated one PIV5 strain from diarrheic piglets. After four times plaque purification and ultracentrifugation, the paramyxovirus‐like particles were observed by electron microscopy. The genome‐wide phylogenetic analysis showed that the isolated strain was closely related to the PIV5 strain from a lesser panda and pigs in China. Therefore, we characterized this isolated PIV5 and found that this virus could haemagglutinate red blood cells from both guinea pigs and chickens. Further, we observed that this PIV5 could infect cell lines from various host species including pig, human, monkey, bovine, dog, cat, rabbit, hamster and mouse, which was confirmed with the immunofluorescent assay. To evaluate the distribution of PIV5 in the field, we developed an indirect ELISA (iELISA) for the first time to detect the specific antibodies based on recombinant nucleocapsid protein. A total of 530 porcine serum samples were tested and the PIV5‐positive rate was 75.7%. To our knowledge, this is the first report describing the full characterization of PIV5 strain isolated from a diarrheic piglet. The ability of this PIV5 strain to infect a wide range of mammalian cell types indicates that PIV5 can transmit across different species, providing a remarkable insight into potential zoonosis. The virus strain and iELISA developed in this study can be used to investigate the pathogenesis, epidemiology, and zoonotic potential of PIV5.

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“…RAA achieves DNA amplification by employing recombinase UvsX, DNA polymerase and single-stranded DNA binding protein at 35°C–42°C, replacing the traditional thermal cycling process ( Fan et al., 2020 ). With the addition of 6-carboxyfluorescein (FAM)-labeled probes and biotin-labeled primers, double-labeled amplification products can be obtained, making the assay sensitive and specific ( He et al., 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of Burkholderia cepacia complex carrying the 16S rRNA gene in clinical specimens by recombinase-aided amplification

Gan

Tian

et al. 2022

Front. Cell. Infect. Microbiol.

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The Burkholderia cepacia complex (BCC) is a group of opportunistic pathogens, including Burkholderia cepacia, Burkholderia multivorans, Burkholderia vietnamiensis and Burkholderia ambifaria, which can cause severe respiratory tract infections and lead to high mortality rates among humans. The early diagnosis and effective treatment of BCC infection are therefore crucial. In this study, a novel and rapid recombinase-aided amplification (RAA) assay targeting the 16S rRNA gene was developed for BCC detection. The protocol for this RAA assay could be completed in 10 min at 39°C, with a sensitivity of 10 copies per reaction and no cross-reactivity with other pathogens. To characterize the effectiveness of the RAA assay, we further collected 269 clinical samples from patients with bacterial pneumonia. The sensitivity and specificity of the RAA assay were 100% and 98.5%, respectively. Seven BCC-infected patients were detected using the RAA assay, and three BCC strains were isolated from the 269 clinical samples. Our data showed that the prevalence of BCC infection was 2.60%, which is higher than the 1.40% reported in previous studies, suggesting that high sensitivity is vital to BCC detection. We also screened a patient with B. vietnamiensis infection using the RAA assay in clinic, allowing for appropriate treatment to be initiated rapidly. Together, these data indicate that the RAA assay targeting the 16S rRNA gene can be applied for the early and rapid detection of BCC pathogens in patients with an uncharacterized infection who are immunocompromised or have underlying diseases, thereby providing guidance for effective treatment.

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Recombinase-Aided Amplification Coupled with Lateral Flow Dipstick for Efficient and Accurate Detection of Porcine Parvovirus

Cited by 9 publications

References 25 publications

Molecular Surveillance for Bocaparvoviruses and Bufaviruses in the European Hedgehog (Erinaceus europaeus)

Molecular Surveillance for Bocaparvoviruses and Bufaviruses in the European Hedgehog (Erinaceus europaeus)

Characterization of parainfluenza virus 5 from diarrheic piglet highlights its zoonotic potential

Rapid detection of Burkholderia cepacia complex carrying the 16S rRNA gene in clinical specimens by recombinase-aided amplification

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