Cortes Aneg L, Montiel Enrique R, Gimeno Isabel M
Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 4700 Hillsborough Street, Raleigh, NC 27606, USA.
Avian Dis. 2009 Dec;53(4):510-6. doi: 10.1637/8871-041009-Reg.1.
The use of Flinders Technology Associates (FTA) filter cards to quantify Marek's disease virus (MDV) DNA for the diagnosis of Marek's disease (MD) and to monitor MD vaccines was evaluated. Samples of blood (43), solid tumors (14), and feather pulp (FP; 36) collected fresh and in FTA cards were analyzed. MDV DNA load was quantified by real-time PCR. Threshold cycle (Ct) ratios were calculated for each sample by dividing the Ct value of the internal control gene (glyceraldehyde-3-phosphate dehydrogenase) by the Ct value of the MDV gene. Statistically significant correlation (P < 0.05) within Ct ratios was detected between samples collected fresh and in FTA cards by using Pearson's correlation test. Load of serotype 1 MDV DNA was quantified in 24 FP, 14 solid tumor, and 43 blood samples. There was a statistically significant correlation between FP (r = 0.95), solid tumor (r = 0.94), and blood (r = 0.9) samples collected fresh and in FTA cards. Load of serotype 2 MDV DNA was quantified in 17 FP samples, and the correlation between samples collected fresh and in FTA cards was also statistically significant (Pearson's coefficient, r = 0.96); load of serotype 3 MDV DNA was quantified in 36 FP samples, and correlation between samples taken fresh and in FTA cards was also statistically significant (r = 0.84). MDV DNA samples extracted 3 days (t0) and 8 months after collection (t1) were used to evaluate the stability of MDV DNA in archived samples collected in FTA cards. A statistically significant correlation was found for serotype 1 (r = 0.96), serotype 2 (r = 1), and serotype 3 (r = 0.9). The results show that FTA cards are an excellent media to collect, transport, and archive samples for MD diagnosis and to monitor MD vaccines. In addition, FTA cards are widely available, inexpensive, and adequate for the shipment of samples nationally and internationally.
对使用弗林德斯技术协会（FTA）滤卡定量马立克氏病病毒（MDV）DNA以诊断马立克氏病（MD）和监测MD疫苗进行了评估。分析了新鲜采集并保存在FTA卡中的血液样本（43份）、实体瘤样本（14份）和羽毛髓样本（FP；36份）。通过实时PCR对MDV DNA载量进行定量。通过将内参基因（甘油醛-3-磷酸脱氢酶）的Ct值除以MDV基因的Ct值，计算每个样本的阈值循环（Ct）比值。使用Pearson相关检验检测新鲜采集和保存在FTA卡中的样本之间Ct比值的统计学显著相关性（P < 0.05）。对24份FP样本、14份实体瘤样本和43份血液样本中的1型MDV DNA载量进行了定量。新鲜采集和保存在FTA卡中的FP样本（r = 0.95）、实体瘤样本（r = 0.94）和血液样本（r = 0.9）之间存在统计学显著相关性。对17份FP样本中的2型MDV DNA载量进行了定量，新鲜采集和保存在FTA卡中的样本之间的相关性也具有统计学显著性（Pearson系数，r = 0.96）；对36份FP样本中的3型MDV DNA载量进行了定量，新鲜采集和保存在FTA卡中的样本之间的相关性也具有统计学显著性（r = 0.84）。采集后3天（t0）和8个月（t1）提取的MDV DNA样本用于评估保存在FTA卡中的存档样本中MDV DNA的稳定性。发现1型（r = 0.96）、2型（r = 1）和3型（r = 0.9）具有统计学显著相关性。结果表明，FTA卡是用于MD诊断和监测MD疫苗的样本采集、运输和存档的优良介质。此外，FTA卡广泛可得、价格低廉，足以在国内和国际运输样本。
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