Validation and comparison of EvaGreen- and TaqMan-based real-time qRT-PCR for diagnosis of the black queen cell virus in the honey bees.

IF 1.8 3区农林科学 Q2 VETERINARY SCIENCES

Veterinary Research Communications Pub Date : 2025-03-06 DOI:10.1007/s11259-025-10706-y

Mustafa Emin Oz, Oguzhan Avci

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引用次数: 0

Abstract

The black queen cell virus (BQCV) is a common agent that causes covert infection in hives and has a global distribution. In this study, TaqMan probe and EvaGreen (EG) dye-based real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays based on the amplification of coding senquences for the BQCV structural protein (SP) and the non-structural protein (NSP) were developed and validated for the detection of BQCV in adult bee and larvae/pupae. Additionally, the performances of commonly used EG and TaqMan chemicals for real-time qRT-PCR analyses were compared. The operating range for BQCV-SP TaqMan and EG real-time qRT-PCRs ranged 2.61-7.61 log₁₀ RNA copies/reaction and 2.61-7.79 log₁₀ RNA copies/reaction, respectively. The operating range for BQCV-NSP TaqMan and EG real-time qRT-PCRs ranged 6.83-11.83 log₁₀ RNA copies/reaction and 6.98-11.98 log₁₀ RNA copies/reaction, respectively. Based on these novel assays, the prevalence of BQCV in adult bees and larvae/pupae was 84% (88/105) and 44% (46/105), respectively. BQCV viral load was also within the operating range determined during assay validation. Comparable results were obtained in analytical and diagnostic performance analyses of the EG dye and TaqMan probe chemicals. Highly sensitive and analytically specific for the detection of BQCV, these real-time qRT-PCR assays will benefit the etiological and epidemiological studies of BQCV.

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基于evgreen和taqman的实时荧光定量pcr诊断蜜蜂黑后细胞病毒的验证与比较

黑后细胞病毒（BQCV）是一种常见的病原体，可引起荨麻疹的隐蔽感染，并具有全球分布。本研究建立了基于TaqMan探针和EvaGreen （EG）染料的BQCV结构蛋白（SP）和非结构蛋白（NSP）编码序列扩增的实时定量反转录聚合酶链反应（qRT-PCR）检测方法，并验证了该方法在成蜂和幼虫/蛹中检测BQCV的有效性。此外，比较了常用的EG和TaqMan化学物质用于实时qRT-PCR分析的性能。BQCV-SP TaqMan和EG实时qrt - pcr的工作范围分别为2.61 ~ 7.61 log10 RNA拷贝/反应和2.61 ~ 7.79 log10 RNA拷贝/反应。BQCV-NSP TaqMan和EG实时qrt - pcr的工作范围分别为6.83 ~ 11.83 log10 RNA拷贝/反应和6.98 ~ 11.98 log10 RNA拷贝/反应。结果表明，BQCV在成蜂和幼虫/蛹中的流行率分别为84%（88/105）和44%（46/105）。BQCV病毒载量也在测定验证时确定的操作范围内。在EG染料和TaqMan探针化学品的分析和诊断性能分析中获得了可比的结果。该方法对BQCV检测具有较高的灵敏度和分析特异性，将有助于BQCV的病原学和流行病学研究。

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来源期刊

Veterinary Research Communications 农林科学-兽医学

CiteScore

2.50

自引率

0.00%

发文量

173

审稿时长

3 months

期刊介绍： Veterinary Research Communications publishes fully refereed research articles and topical reviews on all aspects of the veterinary sciences. Interdisciplinary articles are particularly encouraged, as are well argued reviews, even if they are somewhat controversial. The journal is an appropriate medium in which to publish new methods, newly described diseases and new pathological findings, as these are applied to animals. The material should be of international rather than local interest. As it deliberately seeks a wide coverage, Veterinary Research Communications provides its readers with a means of keeping abreast of current developments in the entire field of veterinary science.