IFA检测石蜡和冰冻切片中鸭病毒性肿头出血症病毒方法建立及在检测病毒鸭体内侵染规律的应用
|
摘要
本文开展了间接免疫荧光(Indirect Immunofluorescence Assay，IFA)检测石蜡切片与冰冻切片中鸭病毒性肿头出血症病毒(Duck Swollenhead HaemorrhagicDisease Virus，DSHDV)方法建立，应用IFA检测DSHDV在实验感染鸭体内侵染规律等系列研究，获如下结果：
1 IFA检测石蜡切片与冰冻切片中鸭病毒性肿头出血症病毒方法建立及其比较
用差速离心纯化DSHDV，将纯化DSHDV免疫家兔制备兔抗DSHDV高免血清，并以High Q阴离子柱层析纯化出兔抗DSHDV IgG，建立间接免疫荧光(IFA)检测石蜡切片与冰冻切片中DSHDV的方法；利用建立的IFA对28日龄鸭人工感染DSHDV死亡鸭不同组织器官以进行检测。IFA检测DSHDV感染死亡鸭石蜡切片肝脏为阳性，检测鸭瘟病毒、禽流感病毒(H5N1)、鸭病毒性肝炎病毒、鸭疫里默氏杆菌感染死亡鸭肝脏呈阴性；IFA检测28日龄人工感染DSHDV死亡鸭的不同组织器官石蜡切片，心、肝、脾、胰、肺、肾、法氏囊、食管、气管、胸肌、胸腺、十二指肠、空肠、回肠、盲肠和直肠为阳性或强阳性；该法具有直观、特异性强的优点，应用于DSHDV在感染鸭组织细胞中的亚细胞定位具有良好效果，可用于DSHDV感染的实验室诊断、病原在鸭组织细胞中分布研究。IFA检测28日龄人工感染DSHDV死亡鸭的不同组织器官冰冻切片，心、肝、脾、胰、肺、肾、法氏囊、食管、气管、胸肌、胸腺、哈德氏腺、十二指肠、空肠、回肠、盲肠和直肠为阳性或强阳性；该法敏感性高于石蜡切片且能快速出结果，可用于临床快速诊断研究。
2 DSHDV在实验感染鸭体内侵染规律的研究
2．1 DSHDV实验感染鸭临床病理学发展规律
采用肌注、口服和滴鼻三种途径感染28日龄北京鸭。感染鸭发病症状：病鸭初期精神沉郁，不愿走动，随着病程的加剧，卧地不起，食欲减退，被毛凌乱，个别鸭流血泪，鼻腔流血性分泌物(90％)。并开始出现特征性症状：头部肿胀(100％)，眼结膜充血、出血呈红色，排灰白色或草绿色稀粪甚至血便。眼观病理变化：心冠脂肪有少量出血斑。头部颈部下黄色胶冻样水肿(80％)。胸腺出血严重，表面和切面有大小不等坏死点(70％)。肝脏呈橙黄色或土黄色，表面有大小不等的出血斑(85％)。心包有淡黄色积液(50％)。肾脏弥漫性出血。气管环状出血，滴鼻组多见(70％)。法氏囊严重出血。食道弥漫性出血形成条状出血带，口服组多见。泄殖腔黏膜初期有出血，后期坏死(50％)。脾肿大且呈深黑色，其上有白色坏死灶(50％)。
2．2 DSHDV在实验感染鸭体内侵染规律
2．2．1肌注组侵染规律
经肌注感染90min后最早能从肾脏中检出病毒抗原；感染后6d仅不能从皮肤、脑、气管、哈德氏腺和腺胃中检测出抗原，该时间段也是病毒达到分布的高峰时期；从不同的时间的检测情况可以看出，经肌注感染病毒在鸭体内分布最为广泛的时间段为感染后12h～6d。
2．2．2口服组侵染规律
DSHDV经口服途径感染初期肾脏、肠道、胸腺、法氏囊的阳性信号较强，而肺脏、心脏阳性信号弱。感染中期大多数组织都检测到了阳性信号，其中肾脏、肠道阳性信号有所增强，脾脏、肌肉、气管开始检测到阳性；哈德氏腺、脑、皮肤、腺胃在感染的整个过程都无阳性信号出现。
2．2．3滴鼻组侵染规律
DSHDV经滴鼻感染后4h最早能从肾脏肾小管上皮细胞浆中检出病毒抗原；感染后12h，能够从肝脏、肾脏、十二指肠和回肠中检出病毒抗原：感染后3d，仅不能从脑、腺胃、皮肤、哈德氏腺中检测出抗原。所以经过滴鼻途径感染DSHDV后24h～6d时间段是病毒达到分布的高峰时期；从不同的时间的检测情况可以看出，经滴鼻感染病毒在鸭体内分布最为广泛的时间段为感染后3d～6d。
综上所述，鸭胚是DSHDV增殖的良好宿主平台，运用建立的IFA对人工感染DSHDV后石蜡切片与冰冻切片中病毒在鸭体内的分布规律进行研究表明：IFA检测28日龄鸭人工感染DSHDV死亡鸭的心、肝、脾、胰、肺、肾、法氏囊、食管、气管、胸肌、胸腺、哈德氏腺、十二指肠、空肠、回肠、盲肠和直肠为阳性；脑、皮肤、腺胃为阴性；阳性组织中病毒抗原主要分布在心脏心肌膜心肌纤维的肌浆中，肝脏肝小叶间的结缔组织和肝索区，胰腺外分泌部腺泡上皮细胞和腺泡心细胞的胞浆中，肾脏肾小管上皮细胞浆，法氏囊粘膜上皮细胞浆，食管黏膜肌层，十二指肠、空肠、直肠、回肠、盲肠肠绒毛上皮细胞浆、肠腺细胞浆中。肠道、肾脏、法氏囊和胸腺是DSHDV侵害的主要靶器官。
This article has established IFA for the Detection of DSHDV in Paraffin-embedded Tissues & Frozen Section. The series studies has applied in detecting the infection characteristics of virus in Ducks. Attains following result:
1 A comparative study for DSHDV detection in Paraffin-embedded tissues and frozen section by IFA
DSHDV was purified by differential centrifugation and rabbit anti-DSHDV antibodies were obtained from rabbits be vaccinated with purified viruses. Then the rabbit anti-DSHDV IgG were purified through High-Q chromotography. IFA was established and used to detect DSHDV antigen in paraffin-embedded tissues of 28-day-old death ducks which experimentally infected with DSHDV. The results were positive in dead ducks infected with DSHDV and negative in dead ducks infected with Duck Plague Virus、Avian influenza virus (H5N1)、Duck viral hepatitis virus and Riemerella Anatipestifer. IFA was also applied in detection of the DSHDV antigen in different organs of artificially infected dead 28 old days ducks, and the antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum and rectum. The establishment of IFA to the detection DSHDV in paraffin-embedded tissues had good specificity, can be applied in diagnosing and studying the distribution of the DSHDV in duck tissues. IFA was also applied in detection of the DSHDV antigen in Frozen Section of artificially infected dead 28 old days ducks, and the viral antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum、harderian gland and rectum. IFA to the detection DSHDV in Frozen Section's sensitivity is higher than in the paraffin-embedded tissues can be applied in clinical diagnosing.
2 The study of the Infection Characteristics of DSHDV in Ducks
2.1 The law of development of clinicopathologic by DSHDV infected
28-day-old beijing ducks which experimentally infected with DSHDV by inject、oral、intranasally. The infected ducks Symptom: Initial period ducks spirit strongly fragrant, not willing to take a walk, Along with course of an illness aggravating, Lies does not get up, anorexia, feather disorderly, some ducks flows tears of blood, nose flows blood secretion (90%). Starts to appear characteristic value symptom: swollen head (100%), conjunctiva hyperaemia or hemorrage, diarrhea with green dejecta even blood excrement. Observations of pathology transformations: Heart crown fat had hemorrhages, frozen type secretion fluid from Subcutaneous (80%). Thymus haemorrhage and the surface and the section had necrosis spots (70%). Yellow and swollen liver with haemorrhage (85%). Heart had yellow accumulates fluid (50%). kidney diffused bleeds. Trachea ring bleeding sees in intranasally (70%). Bursa of fabricius had bleeding. Esophagus diffused hemorrhage forms the strip shape hemorrhage sees in oral. Cloaca mucosal bleed, later ecrosis. Splenomegaly pesents duck blank, above had massive white necrosis stove (50%).
2.2 The Infection Characteristics of DSHDV in Ducks
2.2.1 The Infection Characteristics in inject group
DSHDV can be detected from kidney 90mpi by injected; DSHDV can be detedted from all tissues excepte for skin、brain、trachea、harderian gland and proventriculus at 6dpi, This time section also is the virus achieves the distribution peak time. from the different time detection situation we can see the distribution rate of inject inoculation beteen 12h～6dpi.
2.2.2 The Infection Characteristics in oral group
DSHDV infected by orally, kidney、Intestines、bursa of fabricius、thymus had strong positive, lung、heart had weak positive. Infection intermediate stage, most of tissues were detected positive, in among kidney and intestines had strengthens positive, spleen、muscle、trachea begins detected positive. During the whole process harderian gland、brain、skin and proventriculus were negative.
2.2.3 The Infection Characteristics in intranasal group
DSHDV can be detected from kidney in the cytoplasm of renal tubular epithelial cells 4hpi; DSHDV can be detedted from liver, kidney, duodenum, ileum 12hpi; DSHDV can't be detected only in brain、proventriculus、skin and harderian gland 3dpi. So the the virus achieves the distribution peak time of intranasally inoculation beteen 24h～6dpi. from the different time detection situation we can see the distribution rate of intranasally inoculation beteen 3d～6dpi.
In summary, The duck embryo is good host platform which DSHDV multiplies, Used establishment of IFA for the Detection distributed rule of DSHDV in Paraffin-embedded Tissues & Frozen Section: IFA was detected of the DSHDV antigen in Paraffin-embedded Tissues & Frozen Section of artificially infected dead 28 old days ducks, and the viral antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum、harderian gland and rectum. brain、skin and proventriculus are negative. DSHDV mainly located in the cytoplasm of myocardial fibroblasts, sinus hepaticus in liver, cytoplasm of centroacinar cells in pancreas, cytoplasm of renal tubular epithelial cells in kidney, cytoplasm of epidermic cell of bursa cloacalis, cytoplasm of intestinal epithelial cells in duodenum、jejunum、ileum、cecum and rectum. The target organ that DSHDV mainly attacked were intestines、kidney、bursa of fabricius and thymus.
1 IFA检测石蜡切片与冰冻切片中鸭病毒性肿头出血症病毒方法建立及其比较
用差速离心纯化DSHDV，将纯化DSHDV免疫家兔制备兔抗DSHDV高免血清，并以High Q阴离子柱层析纯化出兔抗DSHDV IgG，建立间接免疫荧光(IFA)检测石蜡切片与冰冻切片中DSHDV的方法；利用建立的IFA对28日龄鸭人工感染DSHDV死亡鸭不同组织器官以进行检测。IFA检测DSHDV感染死亡鸭石蜡切片肝脏为阳性，检测鸭瘟病毒、禽流感病毒(H5N1)、鸭病毒性肝炎病毒、鸭疫里默氏杆菌感染死亡鸭肝脏呈阴性；IFA检测28日龄人工感染DSHDV死亡鸭的不同组织器官石蜡切片，心、肝、脾、胰、肺、肾、法氏囊、食管、气管、胸肌、胸腺、十二指肠、空肠、回肠、盲肠和直肠为阳性或强阳性；该法具有直观、特异性强的优点，应用于DSHDV在感染鸭组织细胞中的亚细胞定位具有良好效果，可用于DSHDV感染的实验室诊断、病原在鸭组织细胞中分布研究。IFA检测28日龄人工感染DSHDV死亡鸭的不同组织器官冰冻切片，心、肝、脾、胰、肺、肾、法氏囊、食管、气管、胸肌、胸腺、哈德氏腺、十二指肠、空肠、回肠、盲肠和直肠为阳性或强阳性；该法敏感性高于石蜡切片且能快速出结果，可用于临床快速诊断研究。
2 DSHDV在实验感染鸭体内侵染规律的研究
2．1 DSHDV实验感染鸭临床病理学发展规律
采用肌注、口服和滴鼻三种途径感染28日龄北京鸭。感染鸭发病症状：病鸭初期精神沉郁，不愿走动，随着病程的加剧，卧地不起，食欲减退，被毛凌乱，个别鸭流血泪，鼻腔流血性分泌物(90％)。并开始出现特征性症状：头部肿胀(100％)，眼结膜充血、出血呈红色，排灰白色或草绿色稀粪甚至血便。眼观病理变化：心冠脂肪有少量出血斑。头部颈部下黄色胶冻样水肿(80％)。胸腺出血严重，表面和切面有大小不等坏死点(70％)。肝脏呈橙黄色或土黄色，表面有大小不等的出血斑(85％)。心包有淡黄色积液(50％)。肾脏弥漫性出血。气管环状出血，滴鼻组多见(70％)。法氏囊严重出血。食道弥漫性出血形成条状出血带，口服组多见。泄殖腔黏膜初期有出血，后期坏死(50％)。脾肿大且呈深黑色，其上有白色坏死灶(50％)。
2．2 DSHDV在实验感染鸭体内侵染规律
2．2．1肌注组侵染规律
经肌注感染90min后最早能从肾脏中检出病毒抗原；感染后6d仅不能从皮肤、脑、气管、哈德氏腺和腺胃中检测出抗原，该时间段也是病毒达到分布的高峰时期；从不同的时间的检测情况可以看出，经肌注感染病毒在鸭体内分布最为广泛的时间段为感染后12h～6d。
2．2．2口服组侵染规律
DSHDV经口服途径感染初期肾脏、肠道、胸腺、法氏囊的阳性信号较强，而肺脏、心脏阳性信号弱。感染中期大多数组织都检测到了阳性信号，其中肾脏、肠道阳性信号有所增强，脾脏、肌肉、气管开始检测到阳性；哈德氏腺、脑、皮肤、腺胃在感染的整个过程都无阳性信号出现。
2．2．3滴鼻组侵染规律
DSHDV经滴鼻感染后4h最早能从肾脏肾小管上皮细胞浆中检出病毒抗原；感染后12h，能够从肝脏、肾脏、十二指肠和回肠中检出病毒抗原：感染后3d，仅不能从脑、腺胃、皮肤、哈德氏腺中检测出抗原。所以经过滴鼻途径感染DSHDV后24h～6d时间段是病毒达到分布的高峰时期；从不同的时间的检测情况可以看出，经滴鼻感染病毒在鸭体内分布最为广泛的时间段为感染后3d～6d。
综上所述，鸭胚是DSHDV增殖的良好宿主平台，运用建立的IFA对人工感染DSHDV后石蜡切片与冰冻切片中病毒在鸭体内的分布规律进行研究表明：IFA检测28日龄鸭人工感染DSHDV死亡鸭的心、肝、脾、胰、肺、肾、法氏囊、食管、气管、胸肌、胸腺、哈德氏腺、十二指肠、空肠、回肠、盲肠和直肠为阳性；脑、皮肤、腺胃为阴性；阳性组织中病毒抗原主要分布在心脏心肌膜心肌纤维的肌浆中，肝脏肝小叶间的结缔组织和肝索区，胰腺外分泌部腺泡上皮细胞和腺泡心细胞的胞浆中，肾脏肾小管上皮细胞浆，法氏囊粘膜上皮细胞浆，食管黏膜肌层，十二指肠、空肠、直肠、回肠、盲肠肠绒毛上皮细胞浆、肠腺细胞浆中。肠道、肾脏、法氏囊和胸腺是DSHDV侵害的主要靶器官。
This article has established IFA for the Detection of DSHDV in Paraffin-embedded Tissues & Frozen Section. The series studies has applied in detecting the infection characteristics of virus in Ducks. Attains following result:
1 A comparative study for DSHDV detection in Paraffin-embedded tissues and frozen section by IFA
DSHDV was purified by differential centrifugation and rabbit anti-DSHDV antibodies were obtained from rabbits be vaccinated with purified viruses. Then the rabbit anti-DSHDV IgG were purified through High-Q chromotography. IFA was established and used to detect DSHDV antigen in paraffin-embedded tissues of 28-day-old death ducks which experimentally infected with DSHDV. The results were positive in dead ducks infected with DSHDV and negative in dead ducks infected with Duck Plague Virus、Avian influenza virus (H5N1)、Duck viral hepatitis virus and Riemerella Anatipestifer. IFA was also applied in detection of the DSHDV antigen in different organs of artificially infected dead 28 old days ducks, and the antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum and rectum. The establishment of IFA to the detection DSHDV in paraffin-embedded tissues had good specificity, can be applied in diagnosing and studying the distribution of the DSHDV in duck tissues. IFA was also applied in detection of the DSHDV antigen in Frozen Section of artificially infected dead 28 old days ducks, and the viral antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum、harderian gland and rectum. IFA to the detection DSHDV in Frozen Section's sensitivity is higher than in the paraffin-embedded tissues can be applied in clinical diagnosing.
2 The study of the Infection Characteristics of DSHDV in Ducks
2.1 The law of development of clinicopathologic by DSHDV infected
28-day-old beijing ducks which experimentally infected with DSHDV by inject、oral、intranasally. The infected ducks Symptom: Initial period ducks spirit strongly fragrant, not willing to take a walk, Along with course of an illness aggravating, Lies does not get up, anorexia, feather disorderly, some ducks flows tears of blood, nose flows blood secretion (90%). Starts to appear characteristic value symptom: swollen head (100%), conjunctiva hyperaemia or hemorrage, diarrhea with green dejecta even blood excrement. Observations of pathology transformations: Heart crown fat had hemorrhages, frozen type secretion fluid from Subcutaneous (80%). Thymus haemorrhage and the surface and the section had necrosis spots (70%). Yellow and swollen liver with haemorrhage (85%). Heart had yellow accumulates fluid (50%). kidney diffused bleeds. Trachea ring bleeding sees in intranasally (70%). Bursa of fabricius had bleeding. Esophagus diffused hemorrhage forms the strip shape hemorrhage sees in oral. Cloaca mucosal bleed, later ecrosis. Splenomegaly pesents duck blank, above had massive white necrosis stove (50%).
2.2 The Infection Characteristics of DSHDV in Ducks
2.2.1 The Infection Characteristics in inject group
DSHDV can be detected from kidney 90mpi by injected; DSHDV can be detedted from all tissues excepte for skin、brain、trachea、harderian gland and proventriculus at 6dpi, This time section also is the virus achieves the distribution peak time. from the different time detection situation we can see the distribution rate of inject inoculation beteen 12h～6dpi.
2.2.2 The Infection Characteristics in oral group
DSHDV infected by orally, kidney、Intestines、bursa of fabricius、thymus had strong positive, lung、heart had weak positive. Infection intermediate stage, most of tissues were detected positive, in among kidney and intestines had strengthens positive, spleen、muscle、trachea begins detected positive. During the whole process harderian gland、brain、skin and proventriculus were negative.
2.2.3 The Infection Characteristics in intranasal group
DSHDV can be detected from kidney in the cytoplasm of renal tubular epithelial cells 4hpi; DSHDV can be detedted from liver, kidney, duodenum, ileum 12hpi; DSHDV can't be detected only in brain、proventriculus、skin and harderian gland 3dpi. So the the virus achieves the distribution peak time of intranasally inoculation beteen 24h～6dpi. from the different time detection situation we can see the distribution rate of intranasally inoculation beteen 3d～6dpi.
In summary, The duck embryo is good host platform which DSHDV multiplies, Used establishment of IFA for the Detection distributed rule of DSHDV in Paraffin-embedded Tissues & Frozen Section: IFA was detected of the DSHDV antigen in Paraffin-embedded Tissues & Frozen Section of artificially infected dead 28 old days ducks, and the viral antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum、harderian gland and rectum. brain、skin and proventriculus are negative. DSHDV mainly located in the cytoplasm of myocardial fibroblasts, sinus hepaticus in liver, cytoplasm of centroacinar cells in pancreas, cytoplasm of renal tubular epithelial cells in kidney, cytoplasm of epidermic cell of bursa cloacalis, cytoplasm of intestinal epithelial cells in duodenum、jejunum、ileum、cecum and rectum. The target organ that DSHDV mainly attacked were intestines、kidney、bursa of fabricius and thymus.
引文
[1] 程安春，汪铭书，陈孝跃，等．一种新发现的鸭病毒性肿头出血症的研究[J]．中国兽医科技，2003，33(10)：15-22．
[2] Badawi A A, Nassif S A,El Ebiary E A. Use of immunofluorescent technique for testing live attenuated vaccines experimentally contaminated with avian reovirus[J].Assiut Veterinary Medical Joumat, 2005, (51):312-320.
[3] Uchiyama A, Besselsen D G.Detection of Reovirus type 3 by use of fluorogenic nuclease reverse transcriptase polymerase chainreaction[J]. LaboratoryAnimals,2003, (37):352-359.
[4] Rajesh S,Agrawal D K,Chauhan R S. Pathomorphological changes in tissues of stunted chickens experimentally infected with avian reovirus[J]. Indian journal of veterinary pathology,2005,29 (1) :1-3.
[5] Liu H J, Kuo L C, Hu Y C.Development of an ELISA for detection of antibodies to avian reovirus in chickens[J]. Journal of Virological Methods,2002,102 (1):129-138.
[6] Bruhn S, Bruckner L,Ottiger H P. Application of RT-PCR for the detection of avian reovirus contamination in avian viral vaccines[J]. Journal of Virological Methods,2005,123 (1) : 179-186.
[7] Kenichi Y, Masanobu G, Kosuke O. Experimental study of swollen head syndrome (SHS) in chickens: pathogenesis and the role of avian reovirus as a trigger in SHS[J].Chiken disease seminarreport.1998,34 (2) :99-107.
[8] 蔡文琴，王波法主编．实用免疫细胞化学与核酸分子杂交技术[M]．四川科学技术出版社，1994．
[9] Pensaert M B,Halterman E O, Bemstein T.Diagnosis of transmissible gastroenteritis in pigs by means of inmmnofluorescence[J].Can J Comp Med, 1968,32 (1):555-556.
[10] Allan G M,M S McNuhy. A direct immunofluorescence test for the rapid detection of avian influenza virus antigen in tissue impression smears[J].Avian Pathol, 1985,14 (5):449-460.
[11] Goel A K, Tamrakar A K, Kamboj D V. Direct immunofluorescence assay for rapid environmental detection of Vibrio cholerae O1 [J]. Folia Microbiologica, 2005,50 (5) :448-452.
[12] Kitamoto N, Kobayashi T, Kato Y. Preparation of monoclonal antibodies cross-reactive with orthopoxviruses and their application for direct immunofluorescence test[J]. Microbiology and immunology, 2005,49 (3) :219-225.
[13] Ilknur T, Pabuccuogtu U, Akin C. Histopathologic and direct immunofluorescence findings of the papulopustular lesions in Behcet's disease[J]. European Journal of Dermatology, 2006,16 (2) :146-150.
[14] Landry M L, Ferguson D, Wlochowski J. Detection of herpes simplex virus in clinical specimens by cytospin-enhanced direct immunofluorescence[J]. Journal of Clinical Microbiology, 1997,35 (1) :302-304.
[15] Foucault C, Rolain J M, Raoult D. Detection of Bartonella quintana by Direct Immunofluorescence Examination of Blood Smears of a Patient with Acute Trench Fever[J]. Journal of Clinical Microbiology, 2004, 42 (10) :4904-4906.
[16] 刘思国，严训南．间接免疫荧光抗体法对鸡传染性支气管炎病毒进行抗原定位[J]．中国预防兽医学报，2000，9(22)：81-84．
[17] 朱建国，焦新安．单抗介导的间接荧光抗体技术快速诊断鸡传染性支气管炎的研究[J]．上海交通大学学报，1995，25(12)：3-5．
[18] Aeeoceberry I, M Theltier, A Achbarou, et al. Production of immonoclonal antibodies directed against the microsporidium En—terocytozoon bieneusi[J]. Journal of Clinical Microbiology, 1999,37 (2) :4107-4112.
[19] Carbajal D,Morano KA,Morano L D.Indirect irnmunofluorescence microscopy for direct detection of Xylella fastidiosa in xylem sap[J].Current Microbiology,2004,49 (5) :372-375.
[20] Bossuyt X, Frans J, Hendrickx A. 'Detection of anti-SSA antibodies by indirect immunofluorescence[J]. Clinical Chemistry, 2004,50 (12) :2361-2369.
[21] Rolf M, Zinkemagel. Immunology Taught by Viruses[J].Science, 1996,271 (1) :173-178.
[22] Tom E M, Morten H. Clinical immunology[J].British Medical Journal, 1996,31 (2) :1465-1469.
[23] 陶义训．免疫学和免疫学检验[M]．北京：人民卫生出版社，1997，166-172．
[24] Doherty PC,Turner SJ,Webby RG.Influenza and the challenge for immunology[J]. Nature immunology, 2006, 7 (5) :449-455.
[25] Lentz A K, Feezor R J. Principles of immunology[J]. Nutrition in Clinical Practice,2003,18 (6) :451-460.
[26] 杜辉，王树迎．组织切片制作中的点滴经验[J]．解剖学杂志，2004，27(2)：165．
[27] Bohon W K, Mesnard R M.New technique of kidney tissue processing for immunofluorescence microscopy: formol sucrose / gum sucrose / paraffin[J]. Lab Invest,1982,4 (1):206-210.
[28] Hamir A N, Moser G, Wampler T. Use of a single anti-nucleocapsid monoclonal antibody to detect rabies antigen in formalin-fixed, paraffin-embedded tissues[J]. The Veterinary Record, 1996,138 (5) :114-115.
[29] Schulz S, Cabras AD, Kremer M. Species identification of mycobacteria in paraffin-embedded tissues: frequent detection of nontuberculous mycobacteria[J]. Modern pathology,2005,18 (2) :274-282.
[30] 胡孟钧．1058例冰冻切片分析[J]．浙江临床医学，2001，3(2)：105．
[31] Suchetha S, Lorenzo M G, Fernando U G. Mucin Stain on Frozen Sections[J]. A Rapid 3-Minute Method. ArchiveS of Pathology and I aboratory Medicine,1999,123 (5) :378-380.
[32] Connolly S S, DArcy F T, Bredin H C. Value of frozen section analysis with suspected testicular malignancy[J]. Urology, 2006,67 (1) :162-165.
[33] Coffin CM, Spilker K, Zhou H. Frozen section diagnosis in pediatric surgical pathology: a decade's experience in a children's hospital[J].Archives of Pathology&Laboratory Medicine, 2005,129 (12) :1619-1625.
[34] Chevez B P. Frozen section diagnosis and indications in ophthalmic pathology[J]. Archives of Pathology & Laboratory Medicine, 2005,129 (12) :1626-1634.
[35] KhalIIF K, Pereira B, Thomas V A. The Accuracy of Intraoperative Frozen Section Analysis of the Sentinel Lymph Nodes During Breast Cancer Surgery[J].International journal of fertility and women's medicine, 2004, 49 (5) :208-211.
[36] Rakha E, Ramaiah S, McGregor A.Accuracy of frozen section in the diagnosis of liver mass lesions[J].Journal of Clinical Pathology, 2006,59 (4) :352-354.
[37] Raab SS, Tworek JA, Souers R. The value of monitoring frozen section-permanent section correlation data over time[J].Archives of Pathology & Laboratory Medicine, 2006,130 (3) :337-342.
[38] F．奥斯伯，R．布伦持，R．E金斯顿，等著．精编分子生物学实验指南[M]．北京：科学出版社，2000，56-97．
[39] 林菊生，冯作化．现代细胞分子生物学技术[M]．北京：科学出版社，2004．
[40] 赵永芳．生物化学技术原理及其应用(第一版)[M]．武汉：武汉大学出版社，1994．
[41] 刘世新主编．生物组织学技术[M]．科学出版社，2004．
[42] 殷震．动物病毒学[M]．第2版．北京：科学出版社，1997．
[43] 吴宝成，陈家样，姚金水，等．番鸭呼肠孤病毒的分离与鉴定[J]，福建农业大学学报，2001，30(2)：227-230．
[44] 傅继华主编．病毒学实用实验技术(第1版)[M]．山东科学技术出版社，2001．
[45] Grande A,Benavente J. Optimal conditions for the growth, purification and storage of the avian reovirus S 1133 [J].Journal of Virological Methods, 2000,85 (1) :43-54.
[46] 卢圣栋主编．现代分子生物学实验技术(第2版)[M]．中国协和医科大学出版社，1999．
[47] Barkon M L,Virgin H W, Haller B L. Circulating immunoglobuling can play a critical role in clearance of intestinal reovirus infection[J]. Journal of Virology, 1996,70 (2) :1109-1116.
[48] 金伯泉．细胞和分子免疫学实验技术[M]．第4军医大学出版社，2002：1-20．
[49] 陈伯权，吴美英，叶群瑞．几种部分纯化单抗方法的比较[J]，病毒学报，1990，6(2)：122-125．
[50] 玛依拉．阿不都克力木，钟旗，康吉克孜·艾沙，张婕．用辛酸—硫酸铵法快速提纯鸡IgG[J]．新疆农业科学，1999，3(1)：25-30．
[51] Sazakli E, Leotsinidis M,Vantarakis A. Comparative typing of Pseudomonas species isolated from the aquatic environment in Greece by SDS-PAGE and RAPD analysis[J]. Journal of applied microbiology, 2005, 99 (5) :1191-1203.
[52] 许宜为．免疫检测技术[M]．北京：科学出版社，1997．
[53] 周春宇，程安春，汪铭书，等．检测鹅细小病毒的间接免疫荧光法的建立[J]．中国兽医杂志，2006，8(42)：19-21．
[54] Marie L, Landry, David Ferguson. Suboptimal detection of influenza correlation of results with the number of antigen—positive cells detected by cytospin immunofluorescence [J]. Journal of Clinical Microbiology, 2003, 41 (7) : 3407-3409.
[55] Allan G M, McNuhy M S.A direct immunofluoreseence test for the rapid detection of avian influenza virus antigen in tissue impression smears[J].Avian Pathol,1985,14:449-460.
[56] 许屏．荧光和免疫荧光染色技术及应用[M]．北京：人民卫生出版社，2000．
[57] Homburger H A,Cahen Y D,Griffiths J, Jacob G L. Detection of antinuclear antibodies: comparative evaluation of enzyme immunoassay and indirect immunofluorescence methods[J]. Archives of Pathology & Laboratory Medicine, 1998,122 (11) :993-999.
[58] 吴小闲主编：医学实验动物微生物学、寄生虫学监测手册[M]．北京，卫生部医学动管会编辑，1992，83-85．
[59] 龚志锦，詹镕洲．病理组织制片和染色技术[M]．上海：上海科学技术出版社，1994：359．
[60] Song J. Characterization of a new monoclonal antibody to a cell surface antign on colorectal cancer and fetal gut tissues[J].Cancer,1986;57:433
[61] Shiroki R, Nazirrudin B, Hoshinaga K.Human peripheral blood lymphocyte-reconstituted, severe combined immunodeficient mice as a model for porcine islet xenograft rejection in humans[J]. Artificial Organs,1996,20 (8) :878-882.
[62] 洪玉萍．介绍一种快速冰冻切片方法[J]．临床与实验病理学杂志，1999，15(5)：351．
[63] 朱礼国，刘涛．制作冰冻切片的体会及技术改进．中华现代临床医学杂志[M]．1999．
[64] Zuhara B, Joanne M, Peter S. Isolation of reoviruses from Pekin ducks in Australia[J]. ACIAR Proceedings, 2004, (2) :81-83.
[65] Guo Z Y, Giambrone J J, Wu H Z, et al. Safety and efficacy of an experimental reovirus vaccine for in ovo administration[J]. Avian Diseases, 2003,47 (4) : 1423-1428.
[66] Nguyen T K, Nguyen V D, Ho V H. Adaptation of duck plague virus to chicken embryo fibroblast cell culture for vaccine production[J]. ACIAR proceedings, 2004,53 (56) :4-5.
[67] Sandhu T S, Shawky S A. Duck virus enteritis (duck plague ) .In:Saif Y M,Barnes H J, Glisson. Diseases of Poultry[M].Ames:lowa State University Press, 2003,354-363.
[68] 程安春，汪铭书，刘菲，等．PCR在鸭瘟临床诊断和免疫及致病机理研究中的初步应用[J]．病毒学报，2004，20(4)：364-370．
[69] 齐雪峰，杨晓燕，罗薇．利用PCR对鸭瘟和鸭“传染性肿头症”的鉴别诊断[J]．四川畜牧兽医，2003，10(30)：21．
[70] 许屏．荧光和免疫荧光染色技术及应用(第二版)[M]．北京：人民卫生出版社，2000．
[71] 阮幼冰主译．免疫荧光基础-实验新原理及其临床新应用(第二版)[M]．北京：人民卫生出版社，1999．
[72] Vadivelu J, Puthucheary S D. Diagnostic and prognostic value of an immunofluorescent assay for melioidosis [J]. The American Journal of Tropical Medicine and Hygiene, 2000, 62 (2) : 297-300.
[73] Graham D A, Foster J C, German A, Mclaren I E, Adair B M, Merza M. Evaluation of an immunofluorescent antibody test to detect bovine herpesvirus 1-specific Ig[J]. Journal of veterinary diagnostic investigation, 1999, 11 (4) : 324-329.
[74] Werner M, Wasielewski R V, Komminoth P. Antigen retrieval,signal amplification and intensification in immunohistochemistry[J]. Histochemistry and Cell Biology, 1996, 105 (4) : 253-260.
[75] Kahveci Z, Minbay F Z, Noyan S, Cavusoglu. A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed, paraffin embedded tissues[J]. Biotechnic & Histochemistry, 2003, 78 (2) : 119-128.
[76] Battifora H, Kopinski M. The influence of protease difestion and duration of fixation on the immunostaining of keratins[J]. The Journal of Histochemistry and Cytochemistry, 1986, 34 (8):1095-1100.
[77] Hazelbag H M, Broek L J, Dorst E- B, Offerhaus G J, Fleuren G J, Hogendoom P C. Immunostaining of chain-specific keratins on formalin-fixed, paraffin- embedded tissues: a comparison of various antigen retrieval systems using microwave heating and proteolytic pre-treatments[J]. The Journal of Histochemistry and Cytochemist ry, 1995, 43 (4) : 429 - 437.
[78] Inoue N, Mar E, Dollard S C, Pau C, Zheng Q, Pellett P E. New immunofluorescence assays for detection of human herpesvirus 8-Specific antibodies[J]. Clinical and Diagnostic Laboratory Immunology, 2000, 7 (3) : 427-435.
[79] Jenson H B, Grant G M, Ench Y, Heard P, Thomas C A, Hilsenbeck S G, Moyer M P. Immunofluorescence microscopy and flow cytometry characterization of chemical induction of latent epstein-barr virus[J]. Clinical and Diagnostic Laboratory Immunology, 1998, 5 (1) : 91-97.
[80] David M ,Wilson, Cesario B. Improved immunodeetion of nuclear antigens after sodiumelodeeyl sulfate treatment of formaldehyde-fixed cells[J]. J Histoelem Cytoehem, 1999,47 (48) :1095.
[81] R. A. B. drury. E. A. wallington. Carleton' s histological technique[J]. 5thed. NewYork: Oxford university press,1980,11 (1) :145.
[82] Bancroft J D. Histochemical technique 2nd[J]. London: Butterworths,1975,28 (2):90.
[83] 骆新兰．冰冻切片常见问题探讨[J]．临床与实验病理学杂志，1999，15(4)：273．
[84] 胡东升，尹建设，左玲．快速冰冻切片机病理制片方法的应用研究[J]．中华今日医学杂志，2003，3(7)：18-50．
[85] Costas C, Costas J M, Bodelon G, Benavente J. The second open reading frame of the avian reovirus S1 gene encodes a transcription-dependent and CRM1-Independent nucleocytoplasmic shuttling protein[J]. Journal of Virology, 2005, 79 (4): 2141-2150.
[86] Ogert R A, Lee L H, Beemon K L. Avian retroviral RNA element promotes unspliced RNA accumulation in the cytoplasm[J]. Journal of Virology, 1996, 70 (6) : 3834-3843.
[87] 欧巧群，姚英民．轮状病毒肠道外感染发病机制的初步探讨[J]．第一军医大学学报，2001，5(1)：111-117．
[88] 魏强．B组轮状病毒引起乳鼠流行性腹泻的病原学与病理学研究[J]．中华实验和临床病毒学杂志，1994，8(3)：204-206．
[89] 蒋玲艳，韦平，李莉萍，等．鸡传染性贫血病毒与其他禽免疫抑制性疾病病毒的混合感染[J]．中国兽医学报，2006，26(6)：591-593．
[2] Badawi A A, Nassif S A,El Ebiary E A. Use of immunofluorescent technique for testing live attenuated vaccines experimentally contaminated with avian reovirus[J].Assiut Veterinary Medical Joumat, 2005, (51):312-320.
[3] Uchiyama A, Besselsen D G.Detection of Reovirus type 3 by use of fluorogenic nuclease reverse transcriptase polymerase chainreaction[J]. LaboratoryAnimals,2003, (37):352-359.
[4] Rajesh S,Agrawal D K,Chauhan R S. Pathomorphological changes in tissues of stunted chickens experimentally infected with avian reovirus[J]. Indian journal of veterinary pathology,2005,29 (1) :1-3.
[5] Liu H J, Kuo L C, Hu Y C.Development of an ELISA for detection of antibodies to avian reovirus in chickens[J]. Journal of Virological Methods,2002,102 (1):129-138.
[6] Bruhn S, Bruckner L,Ottiger H P. Application of RT-PCR for the detection of avian reovirus contamination in avian viral vaccines[J]. Journal of Virological Methods,2005,123 (1) : 179-186.
[7] Kenichi Y, Masanobu G, Kosuke O. Experimental study of swollen head syndrome (SHS) in chickens: pathogenesis and the role of avian reovirus as a trigger in SHS[J].Chiken disease seminarreport.1998,34 (2) :99-107.
[8] 蔡文琴，王波法主编．实用免疫细胞化学与核酸分子杂交技术[M]．四川科学技术出版社，1994．
[9] Pensaert M B,Halterman E O, Bemstein T.Diagnosis of transmissible gastroenteritis in pigs by means of inmmnofluorescence[J].Can J Comp Med, 1968,32 (1):555-556.
[10] Allan G M,M S McNuhy. A direct immunofluorescence test for the rapid detection of avian influenza virus antigen in tissue impression smears[J].Avian Pathol, 1985,14 (5):449-460.
[11] Goel A K, Tamrakar A K, Kamboj D V. Direct immunofluorescence assay for rapid environmental detection of Vibrio cholerae O1 [J]. Folia Microbiologica, 2005,50 (5) :448-452.
[12] Kitamoto N, Kobayashi T, Kato Y. Preparation of monoclonal antibodies cross-reactive with orthopoxviruses and their application for direct immunofluorescence test[J]. Microbiology and immunology, 2005,49 (3) :219-225.
[13] Ilknur T, Pabuccuogtu U, Akin C. Histopathologic and direct immunofluorescence findings of the papulopustular lesions in Behcet's disease[J]. European Journal of Dermatology, 2006,16 (2) :146-150.
[14] Landry M L, Ferguson D, Wlochowski J. Detection of herpes simplex virus in clinical specimens by cytospin-enhanced direct immunofluorescence[J]. Journal of Clinical Microbiology, 1997,35 (1) :302-304.
[15] Foucault C, Rolain J M, Raoult D. Detection of Bartonella quintana by Direct Immunofluorescence Examination of Blood Smears of a Patient with Acute Trench Fever[J]. Journal of Clinical Microbiology, 2004, 42 (10) :4904-4906.
[16] 刘思国，严训南．间接免疫荧光抗体法对鸡传染性支气管炎病毒进行抗原定位[J]．中国预防兽医学报，2000，9(22)：81-84．
[17] 朱建国，焦新安．单抗介导的间接荧光抗体技术快速诊断鸡传染性支气管炎的研究[J]．上海交通大学学报，1995，25(12)：3-5．
[18] Aeeoceberry I, M Theltier, A Achbarou, et al. Production of immonoclonal antibodies directed against the microsporidium En—terocytozoon bieneusi[J]. Journal of Clinical Microbiology, 1999,37 (2) :4107-4112.
[19] Carbajal D,Morano KA,Morano L D.Indirect irnmunofluorescence microscopy for direct detection of Xylella fastidiosa in xylem sap[J].Current Microbiology,2004,49 (5) :372-375.
[20] Bossuyt X, Frans J, Hendrickx A. 'Detection of anti-SSA antibodies by indirect immunofluorescence[J]. Clinical Chemistry, 2004,50 (12) :2361-2369.
[21] Rolf M, Zinkemagel. Immunology Taught by Viruses[J].Science, 1996,271 (1) :173-178.
[22] Tom E M, Morten H. Clinical immunology[J].British Medical Journal, 1996,31 (2) :1465-1469.
[23] 陶义训．免疫学和免疫学检验[M]．北京：人民卫生出版社，1997，166-172．
[24] Doherty PC,Turner SJ,Webby RG.Influenza and the challenge for immunology[J]. Nature immunology, 2006, 7 (5) :449-455.
[25] Lentz A K, Feezor R J. Principles of immunology[J]. Nutrition in Clinical Practice,2003,18 (6) :451-460.
[26] 杜辉，王树迎．组织切片制作中的点滴经验[J]．解剖学杂志，2004，27(2)：165．
[27] Bohon W K, Mesnard R M.New technique of kidney tissue processing for immunofluorescence microscopy: formol sucrose / gum sucrose / paraffin[J]. Lab Invest,1982,4 (1):206-210.
[28] Hamir A N, Moser G, Wampler T. Use of a single anti-nucleocapsid monoclonal antibody to detect rabies antigen in formalin-fixed, paraffin-embedded tissues[J]. The Veterinary Record, 1996,138 (5) :114-115.
[29] Schulz S, Cabras AD, Kremer M. Species identification of mycobacteria in paraffin-embedded tissues: frequent detection of nontuberculous mycobacteria[J]. Modern pathology,2005,18 (2) :274-282.
[30] 胡孟钧．1058例冰冻切片分析[J]．浙江临床医学，2001，3(2)：105．
[31] Suchetha S, Lorenzo M G, Fernando U G. Mucin Stain on Frozen Sections[J]. A Rapid 3-Minute Method. ArchiveS of Pathology and I aboratory Medicine,1999,123 (5) :378-380.
[32] Connolly S S, DArcy F T, Bredin H C. Value of frozen section analysis with suspected testicular malignancy[J]. Urology, 2006,67 (1) :162-165.
[33] Coffin CM, Spilker K, Zhou H. Frozen section diagnosis in pediatric surgical pathology: a decade's experience in a children's hospital[J].Archives of Pathology&Laboratory Medicine, 2005,129 (12) :1619-1625.
[34] Chevez B P. Frozen section diagnosis and indications in ophthalmic pathology[J]. Archives of Pathology & Laboratory Medicine, 2005,129 (12) :1626-1634.
[35] KhalIIF K, Pereira B, Thomas V A. The Accuracy of Intraoperative Frozen Section Analysis of the Sentinel Lymph Nodes During Breast Cancer Surgery[J].International journal of fertility and women's medicine, 2004, 49 (5) :208-211.
[36] Rakha E, Ramaiah S, McGregor A.Accuracy of frozen section in the diagnosis of liver mass lesions[J].Journal of Clinical Pathology, 2006,59 (4) :352-354.
[37] Raab SS, Tworek JA, Souers R. The value of monitoring frozen section-permanent section correlation data over time[J].Archives of Pathology & Laboratory Medicine, 2006,130 (3) :337-342.
[38] F．奥斯伯，R．布伦持，R．E金斯顿，等著．精编分子生物学实验指南[M]．北京：科学出版社，2000，56-97．
[39] 林菊生，冯作化．现代细胞分子生物学技术[M]．北京：科学出版社，2004．
[40] 赵永芳．生物化学技术原理及其应用(第一版)[M]．武汉：武汉大学出版社，1994．
[41] 刘世新主编．生物组织学技术[M]．科学出版社，2004．
[42] 殷震．动物病毒学[M]．第2版．北京：科学出版社，1997．
[43] 吴宝成，陈家样，姚金水，等．番鸭呼肠孤病毒的分离与鉴定[J]，福建农业大学学报，2001，30(2)：227-230．
[44] 傅继华主编．病毒学实用实验技术(第1版)[M]．山东科学技术出版社，2001．
[45] Grande A,Benavente J. Optimal conditions for the growth, purification and storage of the avian reovirus S 1133 [J].Journal of Virological Methods, 2000,85 (1) :43-54.
[46] 卢圣栋主编．现代分子生物学实验技术(第2版)[M]．中国协和医科大学出版社，1999．
[47] Barkon M L,Virgin H W, Haller B L. Circulating immunoglobuling can play a critical role in clearance of intestinal reovirus infection[J]. Journal of Virology, 1996,70 (2) :1109-1116.
[48] 金伯泉．细胞和分子免疫学实验技术[M]．第4军医大学出版社，2002：1-20．
[49] 陈伯权，吴美英，叶群瑞．几种部分纯化单抗方法的比较[J]，病毒学报，1990，6(2)：122-125．
[50] 玛依拉．阿不都克力木，钟旗，康吉克孜·艾沙，张婕．用辛酸—硫酸铵法快速提纯鸡IgG[J]．新疆农业科学，1999，3(1)：25-30．
[51] Sazakli E, Leotsinidis M,Vantarakis A. Comparative typing of Pseudomonas species isolated from the aquatic environment in Greece by SDS-PAGE and RAPD analysis[J]. Journal of applied microbiology, 2005, 99 (5) :1191-1203.
[52] 许宜为．免疫检测技术[M]．北京：科学出版社，1997．
[53] 周春宇，程安春，汪铭书，等．检测鹅细小病毒的间接免疫荧光法的建立[J]．中国兽医杂志，2006，8(42)：19-21．
[54] Marie L, Landry, David Ferguson. Suboptimal detection of influenza correlation of results with the number of antigen—positive cells detected by cytospin immunofluorescence [J]. Journal of Clinical Microbiology, 2003, 41 (7) : 3407-3409.
[55] Allan G M, McNuhy M S.A direct immunofluoreseence test for the rapid detection of avian influenza virus antigen in tissue impression smears[J].Avian Pathol,1985,14:449-460.
[56] 许屏．荧光和免疫荧光染色技术及应用[M]．北京：人民卫生出版社，2000．
[57] Homburger H A,Cahen Y D,Griffiths J, Jacob G L. Detection of antinuclear antibodies: comparative evaluation of enzyme immunoassay and indirect immunofluorescence methods[J]. Archives of Pathology & Laboratory Medicine, 1998,122 (11) :993-999.
[58] 吴小闲主编：医学实验动物微生物学、寄生虫学监测手册[M]．北京，卫生部医学动管会编辑，1992，83-85．
[59] 龚志锦，詹镕洲．病理组织制片和染色技术[M]．上海：上海科学技术出版社，1994：359．
[60] Song J. Characterization of a new monoclonal antibody to a cell surface antign on colorectal cancer and fetal gut tissues[J].Cancer,1986;57:433
[61] Shiroki R, Nazirrudin B, Hoshinaga K.Human peripheral blood lymphocyte-reconstituted, severe combined immunodeficient mice as a model for porcine islet xenograft rejection in humans[J]. Artificial Organs,1996,20 (8) :878-882.
[62] 洪玉萍．介绍一种快速冰冻切片方法[J]．临床与实验病理学杂志，1999，15(5)：351．
[63] 朱礼国，刘涛．制作冰冻切片的体会及技术改进．中华现代临床医学杂志[M]．1999．
[64] Zuhara B, Joanne M, Peter S. Isolation of reoviruses from Pekin ducks in Australia[J]. ACIAR Proceedings, 2004, (2) :81-83.
[65] Guo Z Y, Giambrone J J, Wu H Z, et al. Safety and efficacy of an experimental reovirus vaccine for in ovo administration[J]. Avian Diseases, 2003,47 (4) : 1423-1428.
[66] Nguyen T K, Nguyen V D, Ho V H. Adaptation of duck plague virus to chicken embryo fibroblast cell culture for vaccine production[J]. ACIAR proceedings, 2004,53 (56) :4-5.
[67] Sandhu T S, Shawky S A. Duck virus enteritis (duck plague ) .In:Saif Y M,Barnes H J, Glisson. Diseases of Poultry[M].Ames:lowa State University Press, 2003,354-363.
[68] 程安春，汪铭书，刘菲，等．PCR在鸭瘟临床诊断和免疫及致病机理研究中的初步应用[J]．病毒学报，2004，20(4)：364-370．
[69] 齐雪峰，杨晓燕，罗薇．利用PCR对鸭瘟和鸭“传染性肿头症”的鉴别诊断[J]．四川畜牧兽医，2003，10(30)：21．
[70] 许屏．荧光和免疫荧光染色技术及应用(第二版)[M]．北京：人民卫生出版社，2000．
[71] 阮幼冰主译．免疫荧光基础-实验新原理及其临床新应用(第二版)[M]．北京：人民卫生出版社，1999．
[72] Vadivelu J, Puthucheary S D. Diagnostic and prognostic value of an immunofluorescent assay for melioidosis [J]. The American Journal of Tropical Medicine and Hygiene, 2000, 62 (2) : 297-300.
[73] Graham D A, Foster J C, German A, Mclaren I E, Adair B M, Merza M. Evaluation of an immunofluorescent antibody test to detect bovine herpesvirus 1-specific Ig[J]. Journal of veterinary diagnostic investigation, 1999, 11 (4) : 324-329.
[74] Werner M, Wasielewski R V, Komminoth P. Antigen retrieval,signal amplification and intensification in immunohistochemistry[J]. Histochemistry and Cell Biology, 1996, 105 (4) : 253-260.
[75] Kahveci Z, Minbay F Z, Noyan S, Cavusoglu. A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed, paraffin embedded tissues[J]. Biotechnic & Histochemistry, 2003, 78 (2) : 119-128.
[76] Battifora H, Kopinski M. The influence of protease difestion and duration of fixation on the immunostaining of keratins[J]. The Journal of Histochemistry and Cytochemistry, 1986, 34 (8):1095-1100.
[77] Hazelbag H M, Broek L J, Dorst E- B, Offerhaus G J, Fleuren G J, Hogendoom P C. Immunostaining of chain-specific keratins on formalin-fixed, paraffin- embedded tissues: a comparison of various antigen retrieval systems using microwave heating and proteolytic pre-treatments[J]. The Journal of Histochemistry and Cytochemist ry, 1995, 43 (4) : 429 - 437.
[78] Inoue N, Mar E, Dollard S C, Pau C, Zheng Q, Pellett P E. New immunofluorescence assays for detection of human herpesvirus 8-Specific antibodies[J]. Clinical and Diagnostic Laboratory Immunology, 2000, 7 (3) : 427-435.
[79] Jenson H B, Grant G M, Ench Y, Heard P, Thomas C A, Hilsenbeck S G, Moyer M P. Immunofluorescence microscopy and flow cytometry characterization of chemical induction of latent epstein-barr virus[J]. Clinical and Diagnostic Laboratory Immunology, 1998, 5 (1) : 91-97.
[80] David M ,Wilson, Cesario B. Improved immunodeetion of nuclear antigens after sodiumelodeeyl sulfate treatment of formaldehyde-fixed cells[J]. J Histoelem Cytoehem, 1999,47 (48) :1095.
[81] R. A. B. drury. E. A. wallington. Carleton' s histological technique[J]. 5thed. NewYork: Oxford university press,1980,11 (1) :145.
[82] Bancroft J D. Histochemical technique 2nd[J]. London: Butterworths,1975,28 (2):90.
[83] 骆新兰．冰冻切片常见问题探讨[J]．临床与实验病理学杂志，1999，15(4)：273．
[84] 胡东升，尹建设，左玲．快速冰冻切片机病理制片方法的应用研究[J]．中华今日医学杂志，2003，3(7)：18-50．
[85] Costas C, Costas J M, Bodelon G, Benavente J. The second open reading frame of the avian reovirus S1 gene encodes a transcription-dependent and CRM1-Independent nucleocytoplasmic shuttling protein[J]. Journal of Virology, 2005, 79 (4): 2141-2150.
[86] Ogert R A, Lee L H, Beemon K L. Avian retroviral RNA element promotes unspliced RNA accumulation in the cytoplasm[J]. Journal of Virology, 1996, 70 (6) : 3834-3843.
[87] 欧巧群，姚英民．轮状病毒肠道外感染发病机制的初步探讨[J]．第一军医大学学报，2001，5(1)：111-117．
[88] 魏强．B组轮状病毒引起乳鼠流行性腹泻的病原学与病理学研究[J]．中华实验和临床病毒学杂志，1994，8(3)：204-206．
[89] 蒋玲艳，韦平，李莉萍，等．鸡传染性贫血病毒与其他禽免疫抑制性疾病病毒的混合感染[J]．中国兽医学报，2006，26(6)：591-593．