Production and characterization of monoclonal antibodies for specific detection of Nosema ceranae and Nosema apis in beehive samples.

IF 3.7 2区医学 Q1 PARASITOLOGY International journal for parasitology Pub Date : 2024-12-03 DOI:10.1016/j.ijpara.2024.11.008

Fernando Izquierdo, Carmen Fernández Vadillo, Soledad Fenoy, Carolina Hurtado-Marcos, Angela Magnet, Mariano Higes, Raquel Martín-Hernández, Carmen Del Aguila

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Abstract

Two microsporidian species infect honeybees worldwide, Nosema apis and Nosema ceranae. Two different clinical patterns are considered: nosemosis type A (N. apis) and nosemosis type C (N. ceranae). Nosemosis type A is characterized in acute forms and nosemosis type C shows no clear outward clinical signs. The development of a rapid and simple tool for Nosema detection could allow beekeepers or veterinarians to carry out diagnostic tests in situ. Currently, PCR and microscopy are expensive techniques that require qualified staff and may not be available in every laboratory. The present study describes the production and characterization of four monoclonal antibodies (mAbs) against N. ceranae and N. apis, and the development of an IFAT. An IFAT using the mAbs was compared with microscopy and PCR for 180 beehive samples. The diagnostic test revealed similar sensitivity and specificity percentages to IFAT (97.79% and 93.18%, respectively) and microscopy (97.79% and 95.45%), considering 100% for the PCR as the 'gold standard'. A mAb (7D2) was patented for its high specificity for N. ceranae. The IFAT using the mAbs is a good alternative to microscopy and PCR in laboratories where PCR is not available for the detection and identification of both Nosema spp.

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蜂箱样品中蜡微虫和apis微虫特异性单克隆抗体的制备和鉴定。

两种微孢子虫感染全世界的蜜蜂：蜜蜂微孢子虫和蜜蜂微孢子虫。两种不同的临床模式被考虑：A型鼻窦病（N. apis）和C型鼻窦病（N. ceranae）。A型鼻涕病的特点是急性型，而C型鼻涕病没有明显的外部临床症状。开发一种快速而简单的小虫病检测工具可以让养蜂人或兽医在现场进行诊断测试。目前，聚合酶链反应和显微镜是昂贵的技术，需要合格的工作人员，可能不是每个实验室都有。本研究描述了四种针对蜜蜂和蜜蜂的单克隆抗体（mab）的生产和鉴定，以及IFAT的开发。对180个蜂箱样品的IFAT与显微镜和PCR进行比较。该诊断试验显示与IFAT（分别为97.79%和93.18%）和镜检（分别为97.79%和95.45%）的敏感性和特异性相似，考虑到100%的PCR为“金标准”。A mAb （7D2）因其对ceranae的高特异性而获得专利。使用单克隆抗体的IFAT在实验室中是显微镜和PCR的一个很好的替代方法，在这些实验室中，PCR无法用于检测和鉴定这两种微孢子虫。

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来源期刊

International journal for parasitology 医学-寄生虫学

CiteScore

8.40

自引率

2.50%

发文量

审稿时长

23 days

期刊介绍： International Journal for Parasitology offers authors the option to sponsor nonsubscriber access to their articles on Elsevier electronic publishing platforms. For more information please view our Sponsored Articles page. The International Journal for Parasitology publishes the results of original research in all aspects of basic and applied parasitology, including all the fields covered by its Specialist Editors, and ranging from parasites and host-parasite relationships of intrinsic biological interest to those of social and economic importance in human and veterinary medicine and agriculture.