United States Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural and Veterinary Entomology, 1600 SW 23rd Drive, Gainesville, FL 32608, USA.
J Virol Methods. 2009 Dec;162(1-2):276-9. doi: 10.1016/j.jviromet.2009.07.019. Epub 2009 Jul 29.
Multiplex reverse transcription and polymerase chain reaction (PCR) methods were developed to detect Solenopsis invicta viruses -1, -2, and -3 simultaneously in their host, the red imported fire ant, S. invicta. cDNA synthesis was conducted in a single reaction containing an oligonucleotide primer specific for each virus. Multiplex PCR was subsequently conducted with oligonucleotide primer pairs specific for each virus. The method was specific and sensitive, capable of detecting as few as 500 copies of the viral genomes consistently. Specificity was verified by PCR and amplicon sequencing. The method was evaluated against field-collected samples of ant workers from colonies in Argentina (n=135 ant colonies) and the United States (n=172 ant colonies). The prevalence of each virus in fire ant colonies varied considerably from site to site. A number of colonies exhibited multiple virus infections. However, the multiple SINV infection rate was lower than for single infections. Comparison of viral infection prevalence between S. invicta colonies in Argentina and the U.S. showed no statistical differences, regardless of infection category. This method is anticipated to facilitate epidemiological and related studies concerning the S. invicta viruses in fire ants.
建立了同时检测红火蚁（Solenopsis invicta）体内三种索林诺克斯病毒（Solenopsis invicta virus -1、-2 和 -3）的多重反转录聚合酶链反应（PCR）方法。在单个反应中进行 cDNA 合成，该反应包含针对每种病毒的寡核苷酸引物。随后使用针对每种病毒的寡核苷酸引物对进行多重 PCR。该方法具有特异性和敏感性，能够始终如一地检测到低至 500 个拷贝的病毒基因组。通过 PCR 和扩增子测序验证了特异性。该方法针对来自阿根廷（n=135 个蚁群）和美国（n=172 个蚁群）的野外采集的工蚁样本进行了评估。每种病毒在红火蚁蚁群中的流行程度因地点而异。许多蚁群表现出多种病毒感染。然而，多重 SINV 感染率低于单一感染。无论感染类别如何，阿根廷和美国的红火蚁 S. invicta 病毒感染率均无统计学差异。该方法有望促进有关红火蚁中 S. invicta 病毒的流行病学和相关研究。
