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CN104215774B - 一种利用fgf21因子诊断奶牛隐性酮病的方法 - Google Patents

一种利用fgf21因子诊断奶牛隐性酮病的方法 Download PDF

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CN104215774B
CN104215774B CN201410418651.2A CN201410418651A CN104215774B CN 104215774 B CN104215774 B CN 104215774B CN 201410418651 A CN201410418651 A CN 201410418651A CN 104215774 B CN104215774 B CN 104215774B
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fgf21
ketosis
recessive
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CN104215774A (zh
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徐闯
陈媛媛
夏成
张洪友
许秋实
杨威
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Heilongjiang Bayi Agricultural University
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Abstract

一种利用FGF21因子诊断奶牛隐性酮病的方法。FGF21因子与奶牛隐性酮病呈正相关，应用ELISA试剂盒和生化分析仪测定牛血清中FGF21因子浓度以及酮病、隐性酮病时奶牛血清指标,通过统计学分析FGF21与隐性酮病呈明显相关,与临床酮病无相关性；当335ng/ml<FGF21<443ng/ml时可以诊断为奶牛隐性酮病。该方法确定FGF21与奶牛隐性酮病呈正相关，奶牛血清FGF21可以替代BHBA诊断奶牛隐性酮病，因此能够较早的监测出隐性酮病将会减少奶牛养殖业的损失。

Description

一种利用FGF21因子诊断奶牛隐性酮病的方法
技术领域
本发明涉及一种诊断奶牛隐性酮病的方法，特别是一种利用FGF21因子诊断奶牛隐性酮病的方法。
背景技术
奶牛养殖业是我国畜牧业产业支柱之一,但是奶牛酮病给奶牛业带来了较严重经济损失。尤其是奶牛隐性酮病,由于隐性酮病临床症状不明显，这就给诊断和防治带来了极大的困难，因此较早的监测出隐性酮病将会减少奶牛养殖业的损失。
FGF21是成纤维细胞生长因子家族成员之一。最新研究表明，FGF21刺激脂肪细胞摄取葡萄糖，而对其它类型细胞没有这个作用，这种作用实际上是相当于增强了胰岛素活性。肥胖小鼠注射FGF21后，脂肪组织葡萄糖转运蛋白1显著增加。在饮食诱导肥胖、低血糖和高甘油三酯水平情况下，FGF21对糖尿病老鼠具有保护作用。进一步的研究结果表明，动物注射FGF21后，导致能量消耗增加，脂肪利用和脂质排泄加快。且有FGF21可以用来作为诊断脂肪肝的报道。
目前隐性酮病的监测主要是根据BHBA浓度变化，当1.2mmol/L<BHBA<1.6mmol/L时可诊断为奶牛隐性酮病，但是此种方法敏感性较低，容易造成漏诊。因此需要寻找灵敏度更高的因子进行奶牛隐性酮病的监测。
发明内容
本发明提供一种利用FGF21因子诊断奶牛隐性酮病的方法,所述FGF21对奶牛酮病时酮体改变比较敏感,特别是在奶牛隐性酮病阶段，阐明能量代谢特征性指标BHBA与体内FGF21之间的关系，表明FGF21与奶牛隐性酮病呈正相关。
本发明所采用的技术方案是：一种利用FGF21因子诊断奶牛隐性酮病的方法,FGF21因子与奶牛隐性酮病密切相关，应用ELISA试剂盒和生化分析仪测定牛血清中FGF21因子浓度以及酮病、隐性酮病时奶牛血清指标,通过统计学分析FGF21与隐性酮病呈明显相关,与临床酮病无相关性；当335ng/ml<FGF21<443ng/ml时可以诊断为奶牛隐性酮病。
上述方案中的分析血清FGF21代谢指标与临床酮病组、隐性酮病组、健康组主要检测结果,临床酮病组,隐性酮病组和健康组血清FGF21具有统计学意义，P<0.05；健康组,隐性酮病组、临床酮病组比较血清中BHBA具有统计学意义，P<0.05；隐性酮病时血清FGF21因子与BHBA呈正相关，r＝0.659,p＝0.005；临床酮病时血清FGF21因子与BHBA无相关r＝0.005,p＝0.979。
本发明具有的有益效果是：本发明基于上述背景通过对奶牛临床酮病、隐性酮病和健康奶牛血液中FGF21对奶牛酮病时重要中间代谢产物BHBA水平反应，明确隐性酮病时奶牛体内FGF21变化规律，测量牛群样本中的代谢指标及FGF21因子,确定FGF21与奶牛隐性酮病呈正相关，奶牛血清FGF21可以替代BHBA诊断奶牛隐性酮病，当335ng/ml<FGF21<443ng/ml时可以确定奶牛为隐性酮病，因此能够较早的监测出隐性酮病将会减少奶牛养殖业的损失。
具体实施方式
下面结合实验对本发明进一步说明：
动物实验：奶牛均来自黑龙江省密山某牛场,奶牛间年龄、胎次差异无统计学意义,所有奶牛围产期TMR饲喂。BHBA>1.2mmol/L作为诊断奶牛酮病标准,如果奶牛血清中BHBA>1.6mmol/L，且表现明显临床症状，被诊断为临床酮病组；如果1.2mmol/L<BHBA<1.6mmol/L，且没有明显临床症状，被诊断为隐性酮病组；BHBA<1.2mmol/L且没有任何临床症状表现，作为健康组。其中健康30头,隐性30头,临床30头。此外，其中患有乳腺炎，子宫炎，流产，由于药物导致的肝病，发热等疾病的奶牛被排除本试验。
样品采集和处理
所有奶牛均于早晨空腹抽取尾静脉血，不加抗凝剂，将采样试管置于冰盒之中，1h之内被带回实验室，3000g离心15min，分离血清-20℃下保存备用。
指标测定方法：FGF21使用RD牛成纤维细胞因子ELISA试剂盒，其余代谢指标分别在自动生化分析仪上进行测定。
数据分析：所有数据应用IBMSPSS19.0软件进行分析，计量资料用均数以表示,组间均值差异显著性采用单因素方差统计分析，相关性分析采用Pearson直线相关性分析。p<0.05为显著相关。
实验结果：
单因素方差分析90例奶牛的FGF21,血糖,BHBA,NEFA检测结果见表1。
表1奶牛血清主要检测指标情况
Person相关性分析FGF21代谢指标与酮病的相关性分析见表2。血清FGF21与NEFA呈正相关，与BHBA呈正相关。
表2奶牛FGF21与血清主要指标相关性分析
单因素方差分析不同类型酮病FGF21与酮病代谢指标的关系，90例奶牛中分别为临床酮病30例，隐性酮病30例，健康牛30例。3组牛群胎次、年龄相近，血清FGF21代谢指标与奶牛酮病主要检测结果见表3。临床酮病组,隐性酮病组和健康组血清FGF21(P<0.05),具有统计学意义；健康组、隐性酮病组、临床组血清NEFA与BHBA差异比较(P<0.05),具有统计学意义。
表33组奶牛群FGF21及血清主要检测指标的比较
Person直线相关性分析3组牛群的血清FGF21代谢指标与酮病指标的相关性分析见表4。隐性酮病组血清FGF21与BHBA呈正相关；临床酮病组血清FGF21与BHBA无相关性。
表4不同组群FGF21代谢指标与酮病指标的相关性分析
FGF21与隐性奶牛酮病重要检测指标BHBA呈正相关，FGF21可以用来替代BHBA作为诊断奶牛隐性酮病新因子，当335ng/ml<FGF21<443ng/ml时可以诊断为奶牛隐性酮病。

Claims (2)

1.测定FGF21的试剂在制备奶牛隐性酮病诊断试剂中的用途，其特征在于：FGF21因子与奶牛隐性酮病密切相关，应用ELISA试剂盒和生化分析仪测定奶牛血清中FGF21因子浓度以及酮病、隐性酮病时奶牛血清其余代谢指标，通过统计学分析FGF21与隐性酮病呈明显相关,与临床酮病无相关性；当335ng/ml<FGF21<443ng/ml时可以诊断为奶牛隐性酮病。
2.根据权利要求1所述的用途，其特征在于：分析血清FGF21代谢指标与临床酮病组、隐性酮病组、健康组主要检测结果，临床酮病组、隐性酮病组和健康组血清FGF21具有统计学意义，P<0.05；健康组、隐性酮病组、临床酮病组比较，血清中BHBA具有统计学意义，P<0.05；隐性酮病时血清FGF21因子与BHBA呈正相关，r＝0.659,p＝0.005；临床酮病时血清FGF21因子与BHBA无相关，r＝0.005,p＝0.979。
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AU2017317183A1 (en) * 2016-08-22 2019-02-07 Ambrx, Inc. Bovine fibroblast growth factor 21 and ketosis in dairy cattle
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CN113481221A (zh) * 2021-08-27 2021-10-08 黑龙江八一农垦大学 一种与奶牛酮病相关的血液中编码基因sod及其pcr检测试剂盒

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