干旱区生态环境知识资源中心(Arid): Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes

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DOI	10.1186/s12917-023-03709-x
	Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes
	Souza, Jorge Lucas Nascimento; Silva, Fernanda Cavalcante; da Silva, Carlikelly Gleicy; Bomfim, Isabela Maria Fortaleza Neves; de Medeiros, Henrique Rocha; Zaros, Lilian Giotto
通讯作者	Zaros, LG
来源期刊	BMC VETERINARY RESEARCH
EISSN	1746-6148
出版年	2023
卷号	19 期号:1
英文摘要	Analyzing the stability of reference genes already described as universal is an important methodology to lead gene expression analysis because different studies have shown that the expression of universal reference genes may vary between experimental treatments. In this sense, the glyceraldehyde 3-phosphate dehydrogenase (GAPDH), Succinate dehydrogenase complex subunit A (SDHA) and Ribosomal Protein L-19 (RPL-19) reference genes (already described in other studies with sheep from different regions, breeds and infectious agents or in organisms evolutionarily close to sheep) were investigated in the abomasum, small and large intestines of resistant and susceptible crossbred sheep groups to gastrointestinal nematode infections in the Semi-arid region in Northeast of Brazil. The animals were naturally infected to determine the resistance or susceptibility status by counting eggs per gram (EPG) of feces from the gastrointestinal tract after 33 weeks of observations of infection evolution. Relative gene expression was performed by RT-qPCR methodology using Sybr green and relative gene expression stability was tested by different software programs such as REST, BestKeeper, geNorm and Normfinder. Our results showed the susceptible animals had increase in egg counts per gram of feces than resistant animals (p < 0.001), and both groups showed a mixed infection by nematodes of the genus Haemonchus, Trichostrongylus, Oesophagostomum and Trichuris. Furthermore, we show the importance of analyzing different genes in different software programs and the importance to choose ideal reference genes. In this sense, GAPDH was the most stable gene in the abomasum, whereas SDHA was the most stable in the small and large intestines. In addition, we discuss about variables which can interfere in relative expression such as breed, species, climate and tissue. However, utilizing other reference genes already described in other studies with the same and different variables should be performed.
英文关键词	GAPDH Gastrointestinal tract Sheep and RT-qPCR
类型	Article
语种	英语
开放获取类型	gold
收录类别	SCI-E
WOS记录号	WOS:001064981600003
WOS关键词	TIME QUANTITATIVE PCR ; HAEMONCHUS-CONTORTUS ; EXPRESSION ANALYSIS ; RT-PCR ; RESISTANCE ; MECHANISMS ; IVERMECTIN ; SELECTION ; COOPERIA ; HISTORY
WOS类目	Veterinary Sciences
WOS研究方向	Veterinary Sciences
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/395597
推荐引用方式 GB/T 7714	Souza, Jorge Lucas Nascimento,Silva, Fernanda Cavalcante,da Silva, Carlikelly Gleicy,et al. Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes[J],2023,19(1).
APA	Souza, Jorge Lucas Nascimento,Silva, Fernanda Cavalcante,da Silva, Carlikelly Gleicy,Bomfim, Isabela Maria Fortaleza Neves,de Medeiros, Henrique Rocha,&Zaros, Lilian Giotto.(2023).Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes.BMC VETERINARY RESEARCH,19(1).
MLA	Souza, Jorge Lucas Nascimento,et al."Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes".BMC VETERINARY RESEARCH 19.1(2023).