ABSTRACT
Bean pod mottle virus (BPMV) is a destructive virus that causes serious economic losses in many countries every year, highlighting the importance of its effective detection. In this study, we developed a fast reverse transcription-cross-priming amplification (RT-CPA) coupled with lateral flow dipstick (LFD) diagnostic method for BPMV detection. The RT-CPA-LFD assay that targets the coat protein gene of BPMV was highly specific against diagnosing four other common viruses transmitted by soybean seeds, i.e., Southern bean mosaic virus (SBMV), Tomato ringspot virus (ToRSV), Arabis mosaic virus (ArMV), and Tobacco ringspot virus (TRSV). The sensitivities of the real-time fluorescent RT-CPA and the RT-CPA-LFD assay were at least 50 pg/µl and 500 pg/µl, respectively. Despite a compromise in the limit of detection of the RT-CPA method compared with TaqMan-MGB real-time RT-PCR, our results demonstrated a notably better performance in the detection of field samples of BPMV-infested soybean seeds. With the advantages of efficiency and convenience by visual determination, the RT-CPA-LFD assay presents a potential application for the rapid and accurate detection of BPMV in routine tests.
Subject(s)
Comovirus/isolation & purification , Cross-Priming , Glycine max/virology , Nucleic Acid Amplification Techniques/methods , Plant Diseases/virology , Reverse Transcription , Comovirus/genetics , Sensitivity and SpecificityABSTRACT
The cotton mealybug Phenacoccus solenopsis Tinsley is an emerging invasive insect pest. Since its first report as a pest in the United States in 1991, it has invaded and colonized more than 23 countries over the past century. It was first recorded from Pakistan in 2006 and from China in 2008. In this study, we performed field surveys from 2010 to 2012 and obtained mtCOI sequences from specimens across China and Pakistan, then compared them with already available mtCOI sequences from additional Asian and North American countries. Our genetic analysis provides evidence that P. solenopsis should be classified into two groups, one of which is found only in the United States, and the other found only in Asia. The Asian group contains nine unique haplotypes, two of which have invaded and spread across China, Pakistan, India, and Vietnam over the last 4-6 yr. Our genetic analysis also indicates that P. solenopsis has a close relationship with the parasitoid wasp Aenasius bambawalei Hayat, providing preliminary evidence of a congruent spread of this mealybug and its parasitoids across China.
