PDF(Pubmed)
Abstract：
UNASSIGNED: Infectious bovine keratoconjunctivitis (IBK) is a prevalent ocular disease that affects livestock, leading to substantial economic losses due to reduced production and culling of infected animals. Moraxella spp. is common bacterial pathogens that can cause keratoconjunctivitis in livestock. Therefore, rapid and accurate diagnosis is crucial for effective treatment and disease control. This study aimed to develop a multiplex real-time polymerase chain reaction (mRT-PCR) assay for the detection and differentiation of Moraxella bovoculi, Moraxella ovis, and Moraxella bovis.
UNASSIGNED: Three reference strains of Moraxella as positive controls and 36 lacrimal swab samples collected from cattle were used to evaluate the developed mRT-PCR assay DNA extraction that was performed using the RIBO-sorb DNA/RNA extraction kit. Primers and probes were designed using the SpeciesPrimer pipeline. The annealing temperature, primer and probe concentrations, and sensitivity and specificity of the assay were optimized.
UNASSIGNED: An mRT-PCR assay was developed to detect pathogens associated with IBK in cattle on the basis of optimized parameters. The specificity and sensitivity of this assay were confirmed using samples containing individual pathogens (O - M. ovis, B - M. bovis, and BO - M. bovoculi), combinations of two pathogens (O-B, B-BO, and O-BO), and when the DNA of all three pathogens was present in a single reaction (O-B-BO). The analytical sensitivity of mRT-PCR for detecting M. ovis and M. bovoculi DNA was 21 copies or 50 fg per reaction, whereas that for M. bovis was 210 copies or 500 fg per reaction. In addition, this assay has been tested on samples isolated from the affected eyes of cattle in the Akmola region of the Republic of Kazakhstan.
UNASSIGNED: For the first time in the Republic of Kazakhstan, the proposed mRT-PCR assay for the simultaneous detection of three Moraxella spp. pathogens has been developed. This assay exhibits the required specificity and high sensitivity for m RT-PCR, facilitating the timely implementation of effective measures for disease control and the prevention of economic losses. These losses are linked to a reduction in livestock breeding value, a reduction in meat and milk production, a reduction in the reproductive performance of heifers, resulting in fewer offspring, as well as costs related to the treatment of affected animals.
UNASSIGNED: Three reference strains of Moraxella as positive controls and 36 lacrimal swab samples collected from cattle were used to evaluate the developed mRT-PCR assay DNA extraction that was performed using the RIBO-sorb DNA/RNA extraction kit. Primers and probes were designed using the SpeciesPrimer pipeline. The annealing temperature, primer and probe concentrations, and sensitivity and specificity of the assay were optimized.
UNASSIGNED: An mRT-PCR assay was developed to detect pathogens associated with IBK in cattle on the basis of optimized parameters. The specificity and sensitivity of this assay were confirmed using samples containing individual pathogens (O - M. ovis, B - M. bovis, and BO - M. bovoculi), combinations of two pathogens (O-B, B-BO, and O-BO), and when the DNA of all three pathogens was present in a single reaction (O-B-BO). The analytical sensitivity of mRT-PCR for detecting M. ovis and M. bovoculi DNA was 21 copies or 50 fg per reaction, whereas that for M. bovis was 210 copies or 500 fg per reaction. In addition, this assay has been tested on samples isolated from the affected eyes of cattle in the Akmola region of the Republic of Kazakhstan.
UNASSIGNED: For the first time in the Republic of Kazakhstan, the proposed mRT-PCR assay for the simultaneous detection of three Moraxella spp. pathogens has been developed. This assay exhibits the required specificity and high sensitivity for m RT-PCR, facilitating the timely implementation of effective measures for disease control and the prevention of economic losses. These losses are linked to a reduction in livestock breeding value, a reduction in meat and milk production, a reduction in the reproductive performance of heifers, resulting in fewer offspring, as well as costs related to the treatment of affected animals.
摘要：
■传染性牛角膜结膜炎（IBK）是一种影响家畜的常见眼病，由于受感染动物的产量减少和扑杀，导致巨大的经济损失。莫拉氏菌属。是常见的细菌病原体，可导致牲畜角膜结膜炎。因此,快速准确的诊断对于有效的治疗和疾病控制至关重要。本研究的目的是建立一种多重实时聚合酶链反应(mRT-PCR)检测和鉴别的方法。卵黄莫拉氏菌,还有牛Moraxella.
三个作为阳性对照的莫拉氏菌参考菌株和从牛收集的36个泪腺拭子样品用于评估使用RIBO-sorbDNA/RNA提取试剂盒进行的开发的mRT-PCR测定DNA提取。使用SpeciesPrimer管道设计引物和探针。退火温度,引物和探针浓度，优化了检测的灵敏度和特异性。
■开发了一种mRT-PCR测定法，以在优化参数的基础上检测牛体内与IBK相关的病原体。使用含有单个病原体（O-M.ovis，B-M牛，和BO-M.bovoculi)，两种病原体的组合(O-B，B-BO,和O-BO)，并且当所有三种病原体的DNA存在于单个反应中（O-B-BO）时。mRT-PCR检测M.ovis和bovoculiDNA的分析灵敏度为21拷贝或50fg/反应，而对于牛分枝杆菌，每个反应是210个拷贝或500fg。此外,已在哈萨克斯坦共和国阿克莫拉地区从牛的受影响眼睛分离的样品上测试了该测定。
■第一次在哈萨克斯坦共和国，提出了同时检测三种莫拉氏菌的mRT-PCR方法。病原体已经被开发出来。该方法对mRT-PCR具有所需的特异性和高灵敏度，有利于及时实施有效的疾病控制措施和防止经济损失。这些损失与牲畜育种价值的减少有关，肉类和牛奶产量的减少，小母牛的繁殖性能下降，导致更少的后代，以及与受影响动物的治疗有关的费用。
三个作为阳性对照的莫拉氏菌参考菌株和从牛收集的36个泪腺拭子样品用于评估使用RIBO-sorbDNA/RNA提取试剂盒进行的开发的mRT-PCR测定DNA提取。使用SpeciesPrimer管道设计引物和探针。退火温度,引物和探针浓度，优化了检测的灵敏度和特异性。
■开发了一种mRT-PCR测定法，以在优化参数的基础上检测牛体内与IBK相关的病原体。使用含有单个病原体（O-M.ovis，B-M牛，和BO-M.bovoculi)，两种病原体的组合(O-B，B-BO,和O-BO)，并且当所有三种病原体的DNA存在于单个反应中（O-B-BO）时。mRT-PCR检测M.ovis和bovoculiDNA的分析灵敏度为21拷贝或50fg/反应，而对于牛分枝杆菌，每个反应是210个拷贝或500fg。此外,已在哈萨克斯坦共和国阿克莫拉地区从牛的受影响眼睛分离的样品上测试了该测定。
■第一次在哈萨克斯坦共和国，提出了同时检测三种莫拉氏菌的mRT-PCR方法。病原体已经被开发出来。该方法对mRT-PCR具有所需的特异性和高灵敏度，有利于及时实施有效的疾病控制措施和防止经济损失。这些损失与牲畜育种价值的减少有关，肉类和牛奶产量的减少，小母牛的繁殖性能下降，导致更少的后代，以及与受影响动物的治疗有关的费用。
