Phalen D N, Wilson V G, Graham D L
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College STation 77843-4467.
J Clin Microbiol. 1991 May;29(5):1030-7. doi: 10.1128/jcm.29.5.1030-1037.1991.
A polymerase chain reaction assay was developed for detection of budgerigar fledgling disease virus (BFDV). The assay used a single set of primers complementary to sequences located in the putative coding region for the BFDV VP1 gene. The observed amplification product had the expected size of 550 bp and was confirmed to derive from BFDV DNA by its restriction digestion pattern. This assay was specific for BFDV and highly sensitive, being able to detect as few as 20 copies of the virus. By using the polymerase chain reaction, BFDV was detected in adult, nestling, and embryo budgerigar (Melopsitticus undulatus) tissue DNAs and in sera from adult and nestling budgerigars. These results suggest the possibility of persistent infections in adult birds and lend further support to previously described evidence of possible in ovo transmission. BFDV was also detected in chicken embryo fibroblast cell cultures and chicken eggs inoculated with the virus. A 550-bp product with identical restriction enzyme sites was amplified from a suspected polyomavirus isolated from a peach-faced lovebird (Agapornis pesonata) and from tissue DNA from a Hahn's macaw (Ara nobilis) and a sun conure (Aratinga solstitialis) with histological lesions suggestive of polyomavirus infection. These fragments also hybridized with a BFDV-derived probe, proving that they were derived from a polyomavirus very similar, if not identical, to BFDV.
已开发出一种聚合酶链反应检测法用于检测虎皮鹦鹉雏鸟病病毒（BFDV）。该检测法使用了一组与BFDV VP1基因推定编码区域中的序列互补的引物。观察到的扩增产物大小为预期的550 bp，并通过其限制性消化模式证实源自BFDV DNA。该检测法对BFDV具有特异性且高度灵敏，能够检测低至20个病毒拷贝。通过聚合酶链反应，在成年、雏鸟和胚胎虎皮鹦鹉（虎皮鹦鹉）的组织DNA以及成年和雏鸟虎皮鹦鹉的血清中检测到了BFDV。这些结果表明成年鸟类存在持续性感染的可能性，并进一步支持了先前描述的可能的卵内传播证据。在鸡胚成纤维细胞培养物和接种该病毒的鸡蛋中也检测到了BFDV。从一只桃脸牡丹鹦鹉（桃脸牡丹鹦鹉）分离出的一种疑似多瘤病毒以及一只哈恩金刚鹦鹉（绿翅金刚鹦鹉）和一只太阳锥尾鹦鹉（太阳锥尾鹦鹉）具有提示多瘤病毒感染的组织学病变的组织DNA中扩增出了具有相同限制性酶切位点的550 bp产物。这些片段也与BFDV衍生的探针杂交，证明它们源自一种与BFDV非常相似（如果不是完全相同）的多瘤病毒。
