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美国NovaBios基孔肯雅热试剂盒 基孔肯雅热抗体试剂盒

广州健仑生物科技有限公司

本公司专业供应各种进口品牌基孔肯雅热检测试剂盒，包括美国的NovaBios、德国NOVA、广州创仑等CDC品牌。主要包括胶体金、酶免、PCR等方法学。欢迎咨询

基孔肯雅热IgM诊断试剂

基孔肯雅热IgG诊断试剂

基孔肯雅热ELISA检测试剂

基孔肯雅热快速检测试剂

基孔肯雅病毒核酸检测试剂盒（荧光探针PCR）

美国CDC的基孔肯雅病毒诊断试剂——美国的NovaBios

德国CDC使用的基孔肯雅病毒诊断试剂——德国NOVA

美国NovaBios基孔肯雅热试剂盒 基孔肯雅热抗体试剂盒

【预期用途】 
基孔肯雅IgG/IgM抗体ELISA检测试剂盒主要用于定性检测人血清和血浆中抗基孔肯雅病毒的IgG/IgM抗体。 
【实验原理】 
此试剂盒基于ELISA技术。包被板中包被了抗人IgG抗体，如果人血清或血浆中含有IgG时，则会与其特异性结合，洗板将未结合的物质洗去， 然后加入基孔肯雅抗原溶液，洗板洗去未结合的物质，然后加入链霉亲和素和基孔肯雅抗体酶联物。洗板后，加入TMB底物液，颜色变成蓝色，加入终止液终止反应，颜色由蓝色转为黄色，zui后用酶标仪在450nm处读数。 
【试剂组成】 
包被板：12×8可拆卸，包被了抗人IgG抗体，密封在可重封铝箔袋中 
基孔肯雅溶液1：1瓶包含6mL的基孔肯雅抗原溶液，即用，白盖 
基孔肯雅溶液2：1瓶包含6mL的生物素化的基孔肯雅抗体，即用，蓝色，白盖 
基孔肯雅IgM阳性质控：1瓶，1.5mL，黄色，即用，红盖 
基孔肯雅IgM临界质控：1瓶， 2mL，黄色，即用，绿盖 
基孔肯雅IgM阴性质控：1瓶，1.5mL，黄色，即用，蓝盖 
样本稀释液： 1瓶包含100mL的即用缓冲液，用于稀释样本，pH7.2±0.2，黄色，白盖 
洗涤液：1瓶，包含50mL  20倍浓缩的缓冲液，（pH7.2±0.2）用于洗板，白盖 
链霉亲和素结合液：1瓶包含6mL过氧化物酶结合的链霉亲和素，即用，红色，黑盖 
TMB底物液：1瓶包含15mL  TMB，即用，黄盖 
终止液：1瓶包含15mL，即用，内含硫酸，0.2mol/l，红盖 
【需要的设备和材料】              
固定板
封板片 
酶标仪（450/620nm）              
37℃孵箱 
洗瓶或自动洗板机
10~1000μL的移液器
漩涡混匀器 
蒸馏水或去离子水
一次性试管
计时器 
【储存和稳定性】 
试剂在有效期内，储存于2-8℃稳定 
【试剂准备】 
洗涤液的准备 
用双蒸水稀释洗涤液，例子：10ml洗涤液+190ml双蒸水。稀释好的洗涤液在室温下5天内有效。 
【样本的采集和准备】 
这个实验中使用的样本是人血清和血浆，如果实验在样本采集后的5天内进行，则需要储存在2-8℃，否则，必须于-20℃到-70℃深度冻存。如果样本是深度冻存的，在使用前，则需要充分混匀，避免反复冻融。 不推荐使用热灭活的样本 
【样本的稀释】 
将10μL样本跟1ml的样本稀释液混匀，并用漩涡混匀器充分混匀。
【实验步骤】 
在开始试验前，请仔细阅读试验说明。结果的可信度是依赖于严格地按照实验说明来进行的，铺板时zui少留1个孔为空白对照（A1）1个阴性质控孔（B1）2个临界质控孔（C1+D1）1个阳性质控孔（E1）。开始试验前，请将所有试剂都平衡到室温 
1.  吸取50μL的质控品和稀释过的样本到相应的孔中，留A1孔做空白对照孔
2.  封板 
3.  在37±1℃下孵育1小时±5分钟 
4.  当孵育完成时，揭去封板片，弃去反应液，每孔300μL洗涤液，洗板3次，避免溢出。每孔浸泡的时间都必须＞5秒，zui后拍板将残留的液滴都拍去。 
5.  吸取50μL基孔肯雅溶液1到除了空白对照孔的每个孔中，盖板 
6.  在室温孵育30分钟 
7.  重复步骤4 
8.  将基孔肯雅溶液2跟链霉亲和素结合物混匀10分钟 
9.  吸取50μL基孔肯雅溶液2跟链霉亲和素的复合物到除了空白对照孔的每个孔中，盖板。 
10.  室温孵育30分钟
11.  重复步骤4 
12.  吸取100μL的TMB底物液到每个孔中 
13.  避光孵育15分钟（精确） 
14.  加入100μL终止液到每个孔中，与加TMB底物液时的间隔和顺序都必须一样 
15.  用酶标仪在加入终止液后30分钟内与450/620nm处检测 
【检测】 
调整酶标仪，以空白对照孔调零，以450nm处检测所有孔的吸光度值。 
【结果】 
1.  检测生效的条件 
只有以下条件符合，检测的结果才能认为的有效的  
空白对照孔    吸光度值＜0.100  
阴性质控孔    吸光度值＜临界质控  
临界质控孔    吸光度值0.150-1.300  
阳性质控孔    吸光度值＞临界质控 
如果以上条件不符合的，那么试验结果则是无效的，需要重新检测
2.  结果的计算 
临界质控平均吸光度值的计算，例子：吸光度1：0.39；吸光度2：0.37                                   
（0.39+0.37）/2=0.38    
平均吸光度值为0.38 
3.  结果的说明 
样本如果是比临界值高出10%，则认定为阳性， 
样本如果是在临界值上下10%之内，则认定为灰色区（推荐在2-4周之后再次检测新鲜的样本，如果样本仍然是灰色区，可以直接认为是阴性） 
样本如果是比临界值低出10%，则认定为阴性 
4.  结果的单位 
病人样本平均吸光度值×10 = U   
临界值 
例子： 1.216×10 =32U 
0.38 
临界值： 10 U 
灰色区：9-11 U 
阴性： ＜9 U 
阳性： ＞11 U

美国NovaBios

一、病原学
    CHIKV属于披膜病毒科甲病毒属的Semliki forest（SF）抗原复合群。病毒直径约70nm，有包膜，含有3个结构蛋白（衣壳蛋白C、包膜蛋白E1和E2）和4个非结构蛋白（nsP1、nsP2、nsP3和nsP4）。CHIKV的基因组为不分节段的正链RNA，长度约为11～12 kb。病毒基因组编码顺序为5’－NS1－NS2－NS3－NS4－C－E3－E2－E1－3’。通过病毒部分E1基因的系统发生分析可将CHIKV分为3个组：第1组包含了全部西非的分离株，第2组是亚洲分离株，东、中、南部非洲的分离株构成了第3组。
    CHIKV可在Vero、C6/36、BHK-21和HeLa等细胞中繁殖并产生细胞病变。对血细胞如原代淋巴细胞、T淋巴细胞、B淋巴细胞及单核细胞等不敏感。CHIKV可感染非人灵长类、乳鼠等动物。
    CHIKV对理化因素的抵抗力较弱，对酸、热、脂溶剂、去污剂、漂基孔肯雅热、酚、70％酒精和甲醛敏感。
    二、流行病学
    （一）传染源。
    人和非人灵长类动物是CHIKV的主要宿主。急性期患者、隐基孔肯雅热染者和感染病毒的非人灵长类动物是本病的主要传染源。
    1. 患者：基孔肯雅热急性期患者是主要传染源。人患该病时，在发病后2～5天内可产生高滴度病毒血症，有较强的传染性。
    2．隐基孔肯雅热染者：是CHIKV的重要传染源。
    3．非人灵长类动物：在丛林型疫源地内，亦为本病的主要传染源。已证实非洲绿猴、狒狒、红尾猴、黑猩猩、长臂猿、猕猴和蝙蝠可自然或实验感染CHIKV，并能产生病毒血症。
    （二）传播途径。
    埃及伊蚊(Aedes aegypti)和白纹伊蚊(Ae.albopictus) 是本病的主要传播媒介。主要通过感染病毒的伊蚊叮咬而传播。实验室内可能通过气溶胶传播，目前尚无直接人传人的报道。
    （三）人群易感性。
    人对CHIKV普遍易感，感染后可表现为显基孔肯雅热染或隐基孔肯雅热染。
    （四）流行特征。
    1．地区分布：基孔肯雅热主要分布于非洲、南亚和东南亚地区。在非洲主要流行的国家为坦桑尼亚、南非、津巴布韦、扎伊尔、塞内加尔、安哥拉、尼日利亚、乌干达、罗得西亚、科摩罗、毛里求斯、马达加斯加、马约特岛、塞舌尔及法属留尼旺岛等国家和地区。在亚洲有印度、斯里兰卡、缅甸、越南、泰国、老挝、柬埔寨、菲律宾和马来西亚等。2005－2007年本病在印度洋岛屿、印度和东南亚地区广泛流行，导致数百万人患病。
    2．人群分布：任何年龄均可感染发病，但新老疫区有差异。在新疫区或输入性流行区，所有年龄组均可发病；在非洲和东南亚等长期流行地区，儿童发病较多。无性别、职业和种族差异。
    3．季节分布：本病主要流行季节为夏、秋季，热带地区一年四季均可流行。季节分布主要与媒介的活动有关。
    4．输入性：凡有伊蚊存在地区，当伊蚊达到一定密度且自然条件适合时，如有CHIKV传入，就可能引起流行或暴发。
    三、发病机制与病理改变
    （一）发病机制。
    基孔肯雅热的发病机制目前尚不清楚，近年来的研究有如下看法。
    1．病毒直接侵犯：人被感染CHIKV的蚊子叮咬，约２天后即可发病。发病后第1～2天是高病毒血症期，第3～4天病毒载量下降，通常第5天消失。病毒通过其包膜上的E1、E2蛋白与巨噬细胞、上皮细胞、内皮细胞、成纤维细胞、室管壁膜细胞、小脑膜细胞等细胞上的受体结合，然后通过网格蛋白(calthrin)介导的细胞内吞作用进入细胞，并在细胞内复制，导致细胞坏死和凋亡。
    病毒还可通过胎盘感染胎儿，导致基孔肯雅热或胎儿死亡。
    动物实验证明病毒易侵犯新生小鼠的中枢神经系统、肝、脾及结缔组织。
    2．免疫机制：有研究发现，患者病后2～6天血清中一些细胞因子浓度增高，如干扰素g诱导蛋白-10(CXCL-10)、白细胞介素-8(IL-8)、单核细胞化学趋化蛋白-1（MCP-1)和干扰素g诱导的单核因子（MIG/CXCL9)等，而且以CXCL-10增高为主。患者血清中干扰素g、肿瘤坏死因子a及Th2细胞因子，如IL-1b、IL-6、IL-10和IL-12的浓度保持在正常范围。在恢复期，CXCL-10和MCP-1的浓度下降，由于CXCL-10的功能是在细胞免疫反应中对Th1细胞起化学趋化作用，因此病情严重程度及进展可能与其浓度持续在高水平相关。另外，动物实验证明，干扰素a起着主要的抗病毒作用。
    （二）病理改变。
    1．骨骼肌： 主要感染成纤维细胞,在肌外膜检测到大量的病毒，肌束膜和肌内膜有少量的病毒，而且肌外膜可见巨噬细胞浸润；在肌纤维基底层可见小单核细胞。在感染CHIKV的新生小鼠中可见严重的坏死性肌炎，表现为严重的肌纤维坏死、淋巴细胞和单核巨噬细胞浸润。
    ２．关节：关节囊成纤维细胞可见病毒抗原。
    ３. 皮肤：深真皮层的成纤维细胞可见病毒抗原。
    ４．中枢神经系统：小鼠实验显示，脉络丛上皮细胞严重的空泡变性，脉络丛上皮细胞、室管壁膜细胞和小脑膜细胞有大量的病毒，但脑实质及构成血脑屏障的微血管上皮细胞未见明显改变。
    ５．肝脏：免疫标记及透射电镜显示，在病毒感染小鼠的肝窦毛细血管上皮细胞、巨噬细胞和Kupffer细胞可见病毒抗原及出芽。
    ６．脾脏：在红髓中观察到病毒抗原。
    四、临床表现
    本病的潜伏期为2～12天，通常为3～7天。

美国NovaBios

我司还提供其它进口或国产试剂盒：登革热、疟疾、乙脑、寨卡、黄热病、基孔肯雅热、克锥虫病、违禁品滥用、肺炎球菌、军团菌等试剂盒以及日本生研细菌分型诊断血清、德国SiFin诊断血清、丹麦SSI诊断血清等产品。

想了解更多的NovaBios产品及服务请扫描下方二维码：

【公司名称】 广州健仑生物科技有限公司
【市场部】    杨永汉

【】 
【腾讯  】 2042552662
【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期2幢101-103室

EPIDEMIOLOGY
Chikungunya virus often causes large outbreaks with high attack rates, affecting one-third to three-quarters of the population in areas where the virus is circulating. Outbreaks of chikungunya have occurred in Africa, Asia, Europe, and islands in the Indian and Pacific Oceans. In late 2013, the first locally acquired cases of chikungunya were reported in the Americas on islands in the Caribbean. By the end of 2014, more than 1.1 million suspect cases of chikungunya had been reported in the Americas. Since then the virus has continued to circulate and cause disease in the Americas, Southeast Asia, Pacific Islands, and Africa.
Risk to travelers is highest in areas experiencing ongoing epidemics of the disease (for the most updated information see the Travel Health Notices section on the CDC Travelers’ Health website at wwwnc.cdc.gov/travel/notices). Most epidemics occur during the tropical rainy season and abate during the dry season. However, outbreaks in Africa have occurred after periods of drought, where open water containers in close proximity to human habitation served as vector-breeding sites. Risk of infection exists throughout the day, as the primary vector, Ae. aegypti, aggressively bites during the daytime. Ae. aegypti mosquitoes bite indoors or outdoors near a dwelling. They typically breed in domestic containers that hold water, including buckets and flower pots.
Both adults and children can become infected and symptomatic with the disease. From 2010 through 2013, 110 cases of chikungunya were identified or reported among US travelers who predominantly traveled to areas with known ongoing outbreaks. Following the outbreaks in the Americas, however, >3,500 chikungunya cases have been reported from US states through the end of April 2016. Although most were in travelers, a few cases acquired locally in the continental United States were reported in 2014 and 2015. In addition, several US territories (Puerto Rico, US Virgin Islands, and American Samoa) have reported locally acquired cases from 2014–2016.

CLINICAL PRESENTATION
Approximay 3%–28% of people infected with chikungunya virus will remain asymptomatic. For people who develop symptomatic illness, the incubation period is typically 3–7 days (range, 1–12 days). Disease is most often characterized by sudden onset of high fever (temperature typically >102°F [39°C]) and joint pains. Other symptoms may include headache, myalgia, arthritis, conjunctivitis, nausea, vomiting, or a maculopapular rash. Fevers typically last from several days up to 1 week; the fever can be biphasic. Joint symptoms are often severe and can be debilitating. They usually involve multiple joints, typically bilateral and symmetric. They occur most commonly in hands and feet, but they can affect more proximal joints. Rash usually occurs after onset of fever. It typically involves the trunk and extremities but can also include the palms, soles, and face.
Abnormal laboratory findings can include thrombocytopenia, lymphopenia, and elevated creatinine and liver function tests. Rare but serious complications of the disease can occur, including myocarditis, ocular disease (uveitis, retinitis), hepatitis, acute renal disease, severe bullous lesions, and neurologic disease, such as meningoencephalitis, Guillain-Barré syndrome, myelitis, or cranial nerve palsies. Groups identified as having increased risk for more severe disease include neonates exposed intrapartum, adults >65 years of age, and people with underlying medical conditions, such as hypertension, diabetes, or heart disease.
Acute symptoms of chikungunya typically resolve in 7–10 days. Fatalities associated with infection occur but are typically rare and most commonly reported in older adults. Some patients will have a relapse of rheumatologic symptoms such as polyarthralgia, polyarthritis, tenosynovitis, or Raynaud syndrome in the months after acute illness. Studies have reported variable proportions, ranging from 5% to 80%, of patients with persistent joint pains for months or years after their illness.
Pregnant women have symptoms and outcomes similar to those of other people, and most infections that occur during pregnancy will not result in the virus being transmitted to the fetus. However, when intrapartum transmission occurs, it can result in complications for the baby, including neurologic disease, hemorrhagic symptoms, and myocardial disease. There are also rare reports of spontaneous abortions after maternal infection during the first trimester.

DIAGNOSIS
The differential diagnosis of chikungunya virus infection depends on the clinical signs and symptoms as well as where the person was suspected of being infected. Diseases that should be considered in the differential diagnosis include dengue, Zika, malaria, leptospirosis, parvovirus, enterovirus, group A Streptococcus, rubella, measles, adenovirus, postinfectious arthritis, rheumatologic conditions, or alphavirus infections (including Mayaro, Ross River, Barmah Forest, O’nyong-nyong, and Sindbis viruses).
Preliminary diagnosis is based on the patient’s clinical features, places and dates of travel, and activities. Laboratory diagnosis is generally accomplished by testing serum to detect virus, viral nucleic acid, or virus-specific IgM and neutralizing antibodies. During the first week after onset of symptoms, chikungunya can often be diagnosed by performing viral culture or nucleic acid amplification on serum. Virus-specific IgM and neutralizing antibodies normally develop toward the end of the first week of illness. Therefore, to definitively rule out the diagnosis, convalescent-phase samples should be obtained from patients whose acute-phase samples test negative.
Testing for chikungunya virus is performed at CDC, several state health department laboratories, and several commercial laboratories. Health care providers should report suspected chikungunya cases to their state or local health departments to facilitate diagnosis and mitigate the risk of local transmission. Because chikungunya is a nationally notifiable disease, state health departments should report laboratory-confirmed cases to CDC through ArboNET, the national surveillance system for arboviral diseases.