Comparison of primer pairs: Greater degeneracy improves small ruminant lentivirus (SRLVs) detection by seminested PCR

Dolfini, T.; Conrad, L.F.; Flores, Ismael Vemdrame; Ravazzolo, Ana Paula

doi:10.1016/j.smallrumres.2014.10.007

Cited by 8 publications

(8 citation statements)

References 20 publications

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“…Molecular diagnosis by PCR may add diagnostic value to serodiagnosis since seronegative animals may show PCR positive results due to low antibody production [ 12 , 13 ]. New molecular methods are being described focused on the design of universal primers, thereby increasing sensitivity to enable the identification and removal of animals with low viral load in vivo [ 14 , 15 , 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

Accurate Diagnosis of Small Ruminant Lentivirus Infection Is Needed for Selection of Resistant Sheep through TMEM154 E35K Genotyping

Ramírez

Echeverría

Benito³

et al. 2021

Pathogens

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Small ruminant lentiviruses (SRLV) cause an incurable multiorganic disease widely spread in sheep and goats that disturbs animal welfare and production. In the absence of a vaccine, control measures have been traditionally based on early diagnosis and breeding with virus-inactivated colostrum with segregation of seropositive animals. However, antigenic heterogeneity, poor antibody production due to low viral load, and single strain design of most available ELISA, pose a threat to SRLV diagnosis. Genome-wide association studies have described TMEM154 E35K polymorphism as a good genetic marker for selection of resistant animals in some American and European breeds. In this study, a multitargeted serological and virological screening of more than 500 animals from four different breeds (latxa, raza Navarra, assaf, and churra) attending to SRLV infection status was performed. Then, animals were genotyped to characterize TMEM154 E35K polymorphism. ELISA procedures, individually considered, only identified a proportion of the seropositive animals, and PCR detected a fraction of seronegative animals, globally offering different animal classifications according to SRLV infection status. TMEM154 allele frequency differed substantially among breeds and a positive association between seroprevalence and TMEM154 genotype was found only in one breed. Selection based on TMEM154 may be suitable for specific ovine breeds or SRLV strains, however generalization to the whole SRLV genetic spectrum, ovine breeds, or epidemiological situation may need further validation.

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Section: Introductionmentioning

confidence: 99%

Accurate Diagnosis of Small Ruminant Lentivirus Infection Is Needed for Selection of Resistant Sheep through TMEM154 E35K Genotyping

Ramírez

Echeverría

Benito³

et al. 2021

Pathogens

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“…To achieve sufficient specificity, the primers have to be designed for conserved regions of the viral genome, avoiding the env gene which is less conserved among genotypes [ 18 , 90 ]. On the other hand, the problem of virus genetic variability can be mitigated by the use of degenerate primers expanding the detection range and improving the sensitivity of the method [ 80 , 84 , 91 ]. In infected animals, false negative results of PCR could be linked to co-existence of multiple SRLVs mutants in an infected population.…”

Section: Diagnosis Of Small Ruminant Lentiviral Infectionmentioning

confidence: 99%

Serological, Molecular and Culture-Based Diagnosis of Lentiviral Infections in Small Ruminants

Kalogianni

Stavropoulos

Chaintoutis

et al. 2021

Viruses

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Small ruminant lentiviruses (SRLVs) infections lead to chronic diseases and remarkable economic losses undermining health and welfare of animals and the sustainability of farms. Early and definite diagnosis of SRLVs infections is the cornerstone for any control and eradication efforts; however, a “gold standard” test and/or diagnostic protocols with extensive applicability have yet to be developed. The main challenges preventing the development of a universally accepted diagnostic tool with sufficient sensitivity, specificity, and accuracy to be integrated in SRLVs control programs are the genetic variability of SRLVs associated with mutations, recombination, and cross-species transmission and the peculiarities of small ruminants’ humoral immune response regarding late seroconversion, as well as intermittent and epitope-specific antibody production. The objectives of this review paper were to summarize the available serological and molecular assays for the diagnosis of SRLVs, to highlight their diagnostic performance emphasizing on advantages and drawbacks of their application, and to discuss current and future perspectives, challenges, limitations and impacts regarding the development of reliable and efficient tools for the diagnosis of SRLVs infections.

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“…A utilização de peptídeos combinados pode ser usada para diagnóstico sorológico de triagem da infecção, enquanto que o uso de ELISA com apenas um peptídeo seria útil para sorotipagem (REINA et al, 2009b ;SANJOSÉ et al, 2015) 2009). A combinação do exame sorológico e molecular garante a detecção de um maior número de animais infectados (REINA et al, 2009a;MARINHO et al, 2018;MUZ et al, 2013;DOLFINI et al, 2015;GAYO et al 2019). A aplicação do PCR também é importante ao diagnóstico nos estudos de epidemiologia molecular, assim como para a descrição de novas variantes genéticas em determinada área geográfica (RAMÍREZ et al, 2009;LUJÁN et al, 2019;SCHAER et al, 2022).…”

Section: Ensaio De Imunoadsorção Enzimáticaunclassified

>b<Estudo dos diferentes genótipos de lentivírus de pequenos ruminantes no Brasil e sua relação com manifestações clínicas>/b<

Carvalho

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Ao meu Senhor, Pai dos Céus, agradeço e louvo sempre pela graça derramada na minha vida, e por chegar até aqui, pois tudo posso naquele que me fortalece! Á Universidade de São Paulo, gratidão pela acolhida e oportunidade por também fazer parte da sua história.Á minha orientadora, Profa. Dra. Lilian Gregory, pela oportunidade que me foi concedida para trabalhar com a senhora, pela confiança depositada e os inúmeros momentos de aprendizado pessoal e profissional.Aos meus amigos da equipe Profa.

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Comparison of primer pairs: Greater degeneracy improves small ruminant lentivirus (SRLVs) detection by seminested PCR

Cited by 8 publications

References 20 publications

Accurate Diagnosis of Small Ruminant Lentivirus Infection Is Needed for Selection of Resistant Sheep through TMEM154 E35K Genotyping

Accurate Diagnosis of Small Ruminant Lentivirus Infection Is Needed for Selection of Resistant Sheep through TMEM154 E35K Genotyping

Serological, Molecular and Culture-Based Diagnosis of Lentiviral Infections in Small Ruminants

>b<Estudo dos diferentes genótipos de lentivírus de pequenos ruminantes no Brasil e sua relação com manifestações clínicas>/b<

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