Serological and molecular detection of Anaplasma phagocytophilum in Thoroughbred horses from Chilean racecourses

Anaplasma phagocytophilum is the causative agent of equine granulocytic anaplasmosis (EGA). This study aimed to perform serological and molecular surveys of A. phagocytophilum in thoroughbred horses from racecourses in Chile. Additionally, hematological findings related to A. phagocytophilum molecular positivity were addressed, and phylogenetic analysis of selected positive samples was performed. Complete blood count and msp2 gene real-time PCR were performed in 457 thoroughbred horses from three racecourses located in three different cities of Chile (Santiago, Viña del Mar and Concepción). Sera from horses in two racecourses (Santiago and Vina del Mar) were tested by Indirect fluorescent antibody test (IFAT) to detect IgG antibodies against A. phagocytophilum. The occurrence of A. phagocytophilum by real-time PCR was 13.6 % (62/457, 95 % CI: 10.8–16.3 %), with the highest occurrence observed in Santiago (26.5 %), followed by Concepción (9%), and the lowest in Viña del Mar (5%). The overall frequency of IgG antibodies to A. phagocytophilum was 7.9 % (23/290, 95 % CI: 4.8–12.7 %), with 9.9 % in Santiago and 6.5 % in Viña del Mar. Only three animals from Santiago Racecourse were positive in both real-time PCR and serology. PCR-positive horses from Santiago racecourse presented significantly lower hemoglobin, mean corpuscular value (MCV), and mean corpuscular hemoglobin concentration (CHCM), and higher eosinophil counts. Phylogenetic analysis based on the msp2 gene showed that A. phagocytophilum sequences found in the present study were closely related with A. phagocytophilum sequences from the USA and Europe. Anaplasma phagocytophilum DNA is detected for the first time in Chile.     

VIROLOGICAL AND SEROLOGICAL DIAGNOSIS OF RABIES IN BATS FROM AN URBAN AREA IN THE BRAZILIAN AMAZON

The outbreaks of rabies in humans transmitted by Desmodus rotundus in 2004 and 2005, in the northeast of the Brazilian State of Para, eastern Amazon basin, made this a priority area for studies on this zoonosis. Given this, the present study provides data on this phenomenon in an urban context, in order to assess the possible circulation of the classic rabies virus (RABV) among bat species in Capanema, a town in the Amazon basin. Bats were collected, in 2011, with mist nets during the wet and dry seasons. Samples of brain tissue and blood were collected for virological and serological survey, respectively. None of the 153 brain tissue samples analyzed tested positive for RABV infection, but 50.34% (95% CI: 45.67-55.01%) of the serum samples analyzed were seropositive. Artibeus planirostris was the most common species, with a high percentage of seropositive individuals (52.46%, 95% CI: 52.31 52.60%). Statistically, equal proportions of seropositive results were obtained in the rainy and dry seasons (c² = 0.057, d.f. = 1, p = 0.88). Significantly higher proportions of males (55.96%, 95% CI: 48.96-62.96%) and adults (52.37%, 95% CI: 47.35-57.39%) were seropositive. While none of the brain tissue samples tested positive for infection, the high proportion of seropositive specimens indicates that RABV may be widespread in this urban area.     

A novel SARS-CoV-2 IgG line-blot for evaluating discrepant IgG test results – Observations in pre-pandemic and follow-up samples of five patients

To confirm discrepant SARS-CoV-2-IgG results in four standard assays we applied for the first time a prototype of a coronavirus IgG-line-blot which employs antigens from seasonal coronaviruses, SARS-1 and SARS-CoV-2 combined with avidity testing as a confirmatory tool in a follow-up of five cases including pre-pandemic samples.     

The Sheba Medical Center healthcare workers' children's school: can we open schools safely?

ObjectiveThe role of school closure in mitigating coronavirus disease 2019 (COVID-19) transmission has been questioned. In our medical centre, during a 9-week national lockdown, an alternative school was opened for health-care workers' (HCW) children with a small number of children per class and strict symptom surveillance. After lockdown was lifted we screened children and their parents for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology.MethodsWe conducted a cross-sectional study of HCW parents and their children after one teacher contracted COVID-19 following exposure at home and 53 children were exposed, isolated and tested by RT-PCR. We compared families with children attending the alternative school with families whose children who remained at home during the 9-week lockdown. Epidemiological and medical data were collected using a short questionnaire; nasopharyngeal and oropharyngeal swabs were obtained and tested for SARS-CoV-2 by RT-PCR, and blood was collected for SARS-CoV-2 IgA and IgG titres.ResultsA total of 435 children attended the Sheba alternative school. Among the 53 children exposed to the infected teacher, none tested positive by RT-PCR. Of these, 18 children–parent pairs were tested for serology and all were negative. A total of 106/435 (24%) children and their 78 parents were recruited for the cross-sectional study; 70 attended the Sheba school and 36 did not. Approximately 16% of children in either group reported symptoms (11/70 in the school group and 6/36 in the ‘stay home’ group), but SARS-CoV-2 was not detected by PCR in any, and previous exposure, as determined by serological tests, was low and not significantly different between the groups.ConclusionIn an alternative school for children of HCWs, active during COVID-19 national outbreak, we found no evidence of increased infection compared with children that stayed home.     

地中海贫血患者心肌、肝脏铁沉积与血清铁蛋白相关性研究

目的定量评价地中海贫血患者心脏和肝脏铁过载,探讨心脏、肝脏铁沉积与血清铁蛋白(SF)相关性。材料与方法应用3.0 T磁共振扫描基因确诊地中海贫血患者134例(包括中间型73例和重型61例),扫描包括:肝脏冠状面T2WI及横断面T1WI、T2WI、12回波梯度回波(T2*)序列;心脏标准横断面T2WI及两腔位、四腔位及短轴面电影和8回波梯度回波(T2*)成像。测量心肌、肝脏T2*值,所有受试者MRI扫描前1周完成SF检测。采用Spearman秩相关分析心肌、肝脏铁沉积和SF三者间相关性。结果 134例地中海贫血患者心肌T2*、肝脏T2*、SF中位数分别为23.35(1.88~36.17)ms、1.33(0.36~16.39)ms、1235.3(105.1~14673.0)μg/L,心肌-肝脏T2*(r_s=0.324,P=0.000)、心肌T2*-SF(r_s=-0.491,P=0.000)、肝脏T2*-SF(r_s=-0.697,P=0.000)具有一定相关性。73例中间型地中海贫血患者心肌T2*、肝脏T2*、SF中位数分别为26.18(7.09~36.17)ms、1.81(0.37~16.39)ms、622.8(105.1~10807.0)μg/L,心肌-肝脏T2*(r_s=0.059,P=0.619)、心肌T2*-SF(rs=-0.166,P=0.161)无明显相关,但肝脏T2*-SF间中度负相关(r_s=-0.583,P=0.000)。61例重型地中海贫血患者心肌T2*、肝脏T2*、SF中位数分别为18.80(1.88~33.11)ms、0.72(0.36~10.36)ms、3310.0(313.0~14673.0)μg/L,心肌-肝脏T2*(r_s=0.365,P=0.004)、心肌T2*-SF(r_s=-0.359,P=0.004)、肝脏T2*-SF(r_s=-0.707,P=0.000)具有轻中度相关性。结论在一定范围内,地中海贫血患者心铁沉积与肝铁含量、SF具有较低或无相关性,其间相关性可能随病情加重而增加,而肝铁过载则与SF中度负相关。     

结核病血清诊断中HSP65的应用前景分析

目的：通过评价HSP65抗原表达在结核病血清诊断中的效果,分析其应用前景。方法取活动期结核病患者63例纳入观察组,选取体检健康志愿者60例纳入对照组,就HSP65分别与38 kDa、16 kDa、LAM联合检测诊断对照组与观察组结核病。结果Western-blot结果显示,表达产物在相对分子量65 kμ处有一条表达带,在健康人血清阴性中则无表达带;特异度、敏感度与38 kPa基本一致,差异无统计学意义(P>0.05)。结论 HSP65在结核病血清诊断中可作为备选标志物,反应强度与38kDa基本相当,联合诊断有助于提高诊断符合率;HSP65对结核病X线空洞影敏感度较高。     

The use of pseudotypes to study viruses,virus sero-epidemiology and vaccination

The globalization of the world's economies, accompanied by increasing international travel, changing climates, altered human behaviour and demographics is leading to the emergence of different viral diseases, many of which are highly pathogenic and hence are considered of great public and animal health importance. To undertake basic research and therapeutic development, many of these viruses require handling by highly trained staff in BSL-3/4 facilities not readily available to the majority of the global R&D community. In order to circumvent the enhanced biosafety requirement, the development of non-pathogenic, replication-defective pseudotyped viruses is an effective and established solution to permit the study of many aspects of virus biology in a low containment biosafety level (BSL)-1/2 laboratory. Under the spectre of the unfolding Ebola crisis, this timely conference (the second to be organised by the Viral Pseudotype Unit, www.viralpseudotypeunit.info*) discusses the recent advances in pseudotype technology and how it is revolutionizing the study of important human and animal pathogens (human and avian influenza viruses, rabies/lyssaviruses, HIV, Marburg and Ebola viruses). Key topics addressed in this conference include the exploitation of pseudotypes for serology and serosurveillance, immunogenicity testing of current and next-generation vaccines and new pseudotype assay formats (multiplexing, kit development).*The first pseudotype-focused Euroscicon conference organised by the Viral Pseudotype Unit was recently reviewed [1].