【目的】探究沃柑感染黄龙病的田间症状表现及各症状与感染黄龙病的相关性,为种植者尽早鉴别黄龙病树和及时做好黄龙病防控工作提供参考依据。【方法】从广西不同地区沃柑果园采集沃柑黄龙病疑似样品,对样品按症状分类并记录,用试剂盒...【目的】探究沃柑感染黄龙病的田间症状表现及各症状与感染黄龙病的相关性,为种植者尽早鉴别黄龙病树和及时做好黄龙病防控工作提供参考依据。【方法】从广西不同地区沃柑果园采集沃柑黄龙病疑似样品,对样品按症状分类并记录,用试剂盒法提取叶脉和果柱基因组DNA,采用黄龙病亚洲种的16S r DNA特异引物f D1/fD2和OI1/OI2进行巢式PCR(Nested-PCR)扩增,扩增结果在1%琼脂糖凝胶电泳上检测,统计不同症状样品的阳性率。【结果】收集的214份黄龙病疑似样品中,不同症状样品均检出黄龙病亚洲种病原菌,检出阳性率表现为典型红鼻子果=斑驳黄化叶>果蒂稍红果>均匀黄化叶>叶脉黄化叶>僵果>缺锌黄化叶>其他黄化叶>正常叶。各症状类型与感染黄龙病均存在相关性,但存在差异,其中,典型红鼻子果和斑驳黄化叶阳性率均为100.0%,与感染黄龙病相关性最大;其次果蒂稍红果阳性率为90.9%,与感染黄龙病相关性很大;夏、秋梢均匀黄化叶阳性率为46.5%,与感染黄龙病相关性大。沃柑感染黄龙病后症状复杂,大多出现两种以上复合症状,结果树最初症状为出现果蒂稍红果,落果较多,未结果树以夏、秋梢黄化居多,根系容易受损;正常叶阳性率达16.7%,说明沃柑感染黄龙病早期不易显症。【结论】沃柑出现典型红鼻子果和斑驳黄化叶,可田间诊断为黄龙病树;出现果蒂稍红果,并严重落果,基本可田间诊断为黄龙病树;夏、秋梢均匀黄化黄龙病检出率高,但确诊还需与其他疑似症状(叶脉黄化叶、僵果、缺锌黄化叶、其他黄化叶和正常叶)一样通过PCR进一步检测。Nested-PCR检测技术灵敏性高,可作为沃柑感染黄龙病的早期鉴定技术。展开更多
Objective:To study the prevalence of HHV-8 infection in patients undergoing hemodialysis.Methods:In this study,blood samples of 89 patients undergoing hemodialysis were collected.DNA was extracted from peripheral bloo...Objective:To study the prevalence of HHV-8 infection in patients undergoing hemodialysis.Methods:In this study,blood samples of 89 patients undergoing hemodialysis were collected.DNA was extracted from peripheral blood mononuclear cells and HHV-8 DNA was evaluated by nested-PCR.Results:Of total 89 patients,51(57.3%)were males and 38(42.7%)were females.The patients'age ranged from 24 to 90 years and the mean age was(57.5±1.4)years.HHV-8 DNA was found in 9 of 89(10.1%)peripheral blood mononuclear cell samples,8/51(15.7%)in males and 1/38(2.6%)in females(P=0.07).All patients who were positive for HHV8-DNA were more than 50 years old.Conclusions:This study shows high prevalence of HHV-8.Since hemodialysis patients are candidates for kidney transplantation and due to the possibility of HHV8-reactivation and its serious complications in immunocompromised patients,routine screening for detection of the virus should be implemented for all hemodialysis patients.展开更多
Background:Schistosomiasis japonica is a common zoonosis.Domestic animals are the primary source of infection and play an important role in disease transmission.The prevalence and infectivity of this disease in domest...Background:Schistosomiasis japonica is a common zoonosis.Domestic animals are the primary source of infection and play an important role in disease transmission.The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and,for this reason,diagnostics with a higher sensitivity have become increasingly necessary.It was reported that polymerase chain reaction(PCR)-based methods could be used to detect schistosome infection in humans and animals and presented a high sensitivity and specificity.The present study aimed to develop a PCR-based method for detection of Schistosoma japonicum infection in domestic animals.Methods:A specific nested-PCR assay was developed to detect S.japonicum infection in domestic animals via amplification of a 231-bp DNA fragment of retrotransposon SjR2.The developed assay was first used in sera and dry blood filter paper(DBFP)from goats and buffaloes at different time points of infection.Then,78 DBFPs from 39 artificially-infected bovines at 14 and 28 days post-infection and 42 DBFPs from schistosome-negative bovines from the city of Huangshan in the Anhui province were used to evaluate the diagnostic validity.Furthermore,this assay was used to detect S.japonicum infection in domestic animals in Dongzhi and Wangjiang counties.Results:The expected PCR product was detected in eggs and adult worms of S.japonicum and blood samples from S.japonicum-infected goats and water buffaloes,but not from Fasciola and Haemonchus contortus worms.The nested-PCR assay could detect the target S.japonicum DNA in DBFPs from goats and buffaloes after day 3 post-infection.The sensitivity in buffaloes at 14 and 28 days post-infection was 92.30%(36/39)and 100%(39/39),respectively.The specificity was 97.60%(41/42).The positivity rates in Dongzhi and Wangjiang counties were 6.00%and 8.00%in bovines and 22.00%and 16.67%in goats,respectively.The positivity rates in goats in both counties were higher than those in bovines with a significant difference in Dongzhi County but not in Wangjiang County(P<0.05 and P=0.23,respectively).Conclusions:Our results suggest that the developed nested-PCR assay may be used for the diagnosis of S.japonicum infection in domestic animals,and the control of S.japonicum infection in goats should be paid more attention.展开更多
文摘【目的】探究沃柑感染黄龙病的田间症状表现及各症状与感染黄龙病的相关性,为种植者尽早鉴别黄龙病树和及时做好黄龙病防控工作提供参考依据。【方法】从广西不同地区沃柑果园采集沃柑黄龙病疑似样品,对样品按症状分类并记录,用试剂盒法提取叶脉和果柱基因组DNA,采用黄龙病亚洲种的16S r DNA特异引物f D1/fD2和OI1/OI2进行巢式PCR(Nested-PCR)扩增,扩增结果在1%琼脂糖凝胶电泳上检测,统计不同症状样品的阳性率。【结果】收集的214份黄龙病疑似样品中,不同症状样品均检出黄龙病亚洲种病原菌,检出阳性率表现为典型红鼻子果=斑驳黄化叶>果蒂稍红果>均匀黄化叶>叶脉黄化叶>僵果>缺锌黄化叶>其他黄化叶>正常叶。各症状类型与感染黄龙病均存在相关性,但存在差异,其中,典型红鼻子果和斑驳黄化叶阳性率均为100.0%,与感染黄龙病相关性最大;其次果蒂稍红果阳性率为90.9%,与感染黄龙病相关性很大;夏、秋梢均匀黄化叶阳性率为46.5%,与感染黄龙病相关性大。沃柑感染黄龙病后症状复杂,大多出现两种以上复合症状,结果树最初症状为出现果蒂稍红果,落果较多,未结果树以夏、秋梢黄化居多,根系容易受损;正常叶阳性率达16.7%,说明沃柑感染黄龙病早期不易显症。【结论】沃柑出现典型红鼻子果和斑驳黄化叶,可田间诊断为黄龙病树;出现果蒂稍红果,并严重落果,基本可田间诊断为黄龙病树;夏、秋梢均匀黄化黄龙病检出率高,但确诊还需与其他疑似症状(叶脉黄化叶、僵果、缺锌黄化叶、其他黄化叶和正常叶)一样通过PCR进一步检测。Nested-PCR检测技术灵敏性高,可作为沃柑感染黄龙病的早期鉴定技术。
基金This study was financially supported by Student Research Committee,Ahvaz Jundishapur University of Medical Sciences,Ahvaz,Iran.
文摘Objective:To study the prevalence of HHV-8 infection in patients undergoing hemodialysis.Methods:In this study,blood samples of 89 patients undergoing hemodialysis were collected.DNA was extracted from peripheral blood mononuclear cells and HHV-8 DNA was evaluated by nested-PCR.Results:Of total 89 patients,51(57.3%)were males and 38(42.7%)were females.The patients'age ranged from 24 to 90 years and the mean age was(57.5±1.4)years.HHV-8 DNA was found in 9 of 89(10.1%)peripheral blood mononuclear cell samples,8/51(15.7%)in males and 1/38(2.6%)in females(P=0.07).All patients who were positive for HHV8-DNA were more than 50 years old.Conclusions:This study shows high prevalence of HHV-8.Since hemodialysis patients are candidates for kidney transplantation and due to the possibility of HHV8-reactivation and its serious complications in immunocompromised patients,routine screening for detection of the virus should be implemented for all hemodialysis patients.
基金This study was supported by research funds provided by the Special Fund for Agro-scientific Research in the Public Interest(grant No.201303037)the National Natural Science Foundation of China(grant No.31572218)the Scientific and Technological Innovation Program of the Chinese Academy of Agricultural Sciences。
文摘Background:Schistosomiasis japonica is a common zoonosis.Domestic animals are the primary source of infection and play an important role in disease transmission.The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and,for this reason,diagnostics with a higher sensitivity have become increasingly necessary.It was reported that polymerase chain reaction(PCR)-based methods could be used to detect schistosome infection in humans and animals and presented a high sensitivity and specificity.The present study aimed to develop a PCR-based method for detection of Schistosoma japonicum infection in domestic animals.Methods:A specific nested-PCR assay was developed to detect S.japonicum infection in domestic animals via amplification of a 231-bp DNA fragment of retrotransposon SjR2.The developed assay was first used in sera and dry blood filter paper(DBFP)from goats and buffaloes at different time points of infection.Then,78 DBFPs from 39 artificially-infected bovines at 14 and 28 days post-infection and 42 DBFPs from schistosome-negative bovines from the city of Huangshan in the Anhui province were used to evaluate the diagnostic validity.Furthermore,this assay was used to detect S.japonicum infection in domestic animals in Dongzhi and Wangjiang counties.Results:The expected PCR product was detected in eggs and adult worms of S.japonicum and blood samples from S.japonicum-infected goats and water buffaloes,but not from Fasciola and Haemonchus contortus worms.The nested-PCR assay could detect the target S.japonicum DNA in DBFPs from goats and buffaloes after day 3 post-infection.The sensitivity in buffaloes at 14 and 28 days post-infection was 92.30%(36/39)and 100%(39/39),respectively.The specificity was 97.60%(41/42).The positivity rates in Dongzhi and Wangjiang counties were 6.00%and 8.00%in bovines and 22.00%and 16.67%in goats,respectively.The positivity rates in goats in both counties were higher than those in bovines with a significant difference in Dongzhi County but not in Wangjiang County(P<0.05 and P=0.23,respectively).Conclusions:Our results suggest that the developed nested-PCR assay may be used for the diagnosis of S.japonicum infection in domestic animals,and the control of S.japonicum infection in goats should be paid more attention.
