Linnen Jeffrey M, Vinelli Elizabeth, Sabino Ester C, Tobler Leslie H, Hyland Catherine, Lee Tzong-Hae, Kolk Daniel P, Broulik Amy S, Collins Cynthia S, Lanciotti Robert S, Busch Michael P
Gen-Probe, Inc., San Diego, California, USA.
Transfusion. 2008 Jul;48(7):1355-62. doi: 10.1111/j.1537-2995.2008.01772.x. Epub 2008 May 22.
Dengue fever and hemorrhagic disease are caused by four dengue virus (DENV) serotypes (DENV-1 to -4), mosquito-borne flaviviruses with increasing incidence, and expanding global distributions. Documented transfusion transmission of West Nile virus raised concern regarding transfusion-transmitted DENV.
A DENV RNA assay was developed based on transcription-mediated amplification (TMA) blood screening assays routinely used by blood centers worldwide. Sensitivity was established by endpoint dilution analyses of DENV-1 RNA transcript and pedigreed tissue culture standards for all four DENV-serotypes. Frozen plasma samples were tested from 2994 donations from Honduras (September 2004-January 2005), 4858 donations from Brazil (February-April 2003), and 5879 donations from Australia (March-September 2003). Type-specific polymerase chain reaction (PCR) assays were used to quantify and genotype TMA repeat-reactive samples; viral cultures, type-specific antibody, and antigen assays were also performed.
The TMA assay detected 14.9 copies per mL DENV-1 transcript (95% detection limit), with comparable sensitivity for all four serotypes. Honduran donors yielded 9 TMA repeat-reactive samples (0.30%); 8 were confirmed by PCR, with 3 DENV serotypes detected and viral loads from fewer than 3 x 10(4) to 4.2 x 10(4) copies per mL; and 4 samples yielded infectious virus. Three (0.06%) Brazilian samples tested repeat-reactive; 2 (0.04%) were PCR-positive (serotypes DENV-1 and -3; 12 and 294 copies/mL). No Australian donor samples tested repeat-reactive.
Dengue viremia rates among asymptomatic blood donors ranged from 0.30 percent in Honduras to 0.04 percent in Brazil. Future studies are needed to establish rates of transfusion transmission by viremic donations and clinical consequences in recipients.
登革热和出血热由四种登革病毒(DENV)血清型(DENV-1至-4)引起,这些病毒是通过蚊子传播的黄病毒,发病率不断上升,全球分布范围不断扩大。有记录的西尼罗河病毒输血传播引发了对输血传播DENV的担忧。
基于全球血液中心常规使用的转录介导扩增(TMA)血液筛查检测方法开发了一种DENV RNA检测方法。通过对DENV-1 RNA转录本和所有四种DENV血清型的纯种组织培养标准品进行终点稀释分析来确定灵敏度。对来自洪都拉斯(2004年9月至2005年1月)的2994份献血、来自巴西(2003年2月至4月)的4858份献血以及来自澳大利亚(2003年3月至9月)的5879份献血的冷冻血浆样本进行检测。使用型特异性聚合酶链反应(PCR)检测方法对TMA重复反应性样本进行定量和基因分型;还进行了病毒培养、型特异性抗体和抗原检测。
TMA检测方法检测到每毫升DENV-1转录本有14.9个拷贝(95%检测限),对所有四种血清型的灵敏度相当。洪都拉斯的献血者中有9份样本TMA重复反应阳性(0.30%);8份经PCR确认,检测到3种DENV血清型,病毒载量从每毫升少于3×10⁴至4.2×10⁴个拷贝;4份样本产生了感染性病毒。3份(0.06%)巴西样本检测为重复反应阳性;2份(0.04%)PCR呈阳性(血清型为DENV-1和-3;每毫升12和