血清法监测人工感染弓形虫病猪特异抗体的比较

应用ELISA、LAT、IHA血清试验对8头人工感染弓形虫猪的血清抗体滴度变化规律进行了监测,并对不同检测方法的结果进行了比较.结果发现,试验猪在皮下感染弓形虫RH株速殖子后,血清中的特异性抗体首次检出的时间方面,LAT(第4 d)早于ELISA(第14 d)和IHA(第20 d);检出持续时间方面,LAT(54 d)长于ELISA(44 d)和IHA(19 d);平均检出率方面,LAT(7/8)高于ELISA(6.7/8)和IHA(5/8).试验结果表明,ELISA和LAT适合于猪弓形虫病的诊断和血清学调查,LAT更适合于猪弓形虫病的早期检测.     

LAT52启动子在甘蓝花药中的表达模式

[目的]验证花药特异性启动子LAT52能否在甘蓝花药中发挥功能。[方法]从番茄中扩增得到LAT52核心序列608 bp,进行序列分析,连接LAT52启动子到双元表达载体p BI121,转化农杆菌GV3101,用农杆菌浸染法侵染甘蓝幼苗下胚轴,获得2株阳性植株,对转基因甘蓝植株的花蕾、花药及花粉进行GUS染色。[结果]LAT52核心序列608 bp具多个与花粉特异性表达相关的元件,浸染的LAT52启动子仅在花药及花粉中表达出蓝色,其他部位未染上蓝色。[结论]LAT52启动子仅在甘蓝花药及花粉中特异性表达。     

Evidence of LAT1 expression in canine caput epididymis

L-type amino acid transporter 1 (LAT1), the first isotype of amino acid transport system L, transports aromatic and branched amino acids pivotal for fundamental cellular activities such cellular growth and proliferation. LAT1 expression was high only in the brain in contrast to its limited distribution and low level of expression in normal tissues. We found potent LAT1 expression in canine caput epididymis by quantitative RT-PCR and Western blotting analysis. Immnuno-histochemical examination revealed observable LAT1 in microvillous epithelial cells.     

L型氨基酸转运载体1对奶牛乳腺中β-酪蛋白表达的影响

为研究L型氨基酸转运载体1（L type amino acid transporter 1,LAT1）对奶牛乳腺中β-酪蛋白表达的作用,本试验采用PCR技术体外扩增奶牛LAT1基因并构建LAT1真核表达载体,采用脂质体转染技术将LAT1真核表达载体转染奶牛乳腺上皮细胞,并于转染48 h后采用Western blotting技术检测LAT1过表达后乳腺上皮细胞中LAT1、4F2hc和β-酪蛋白的表达变化。荧光显微镜检测结果显示,LAT1真核表达载体成功转染奶牛乳腺上皮细胞;Western blotting检测结果显示,LAT1过表达的奶牛乳腺上皮细胞中LAT1极显著增加（P<0.01）,β-酪蛋白的表达显著升高（P<0.05）,4F2hc表达变化不显著（P>0.05）。这些结果提示LAT1对奶牛乳腺上皮细胞乳蛋白合成具有促进作用。     

乳胶凝集试验与血清中和试验检测猪伪狂犬病血清抗体的 …

应用血清中和试验（ＳＮＴ）和伪狂犬病乳胶凝集试验（ＬＡＴ）诊断试剂盒对两种伪狂犬病是性血清、伪狂犬病病毒（ＰＲＶ）高兔血甭及６０份被检猪血清进行了ＰＲＶ抗体效价测定和相关性分析,两种方法测得的抗体效价之间呈强相关性（ｒ＝０．９６）,且ＬＡＴ效价比ＳＮＴ一般高出一个滴度;能干为自３５个猪场的４１４份猪血清进行了ＰＲＶ抗体检测,并与ＳＮＴ检测结果进行了对比,结果在ＳＮＴ检测为阳笥的１７１份血清中,ＬＡ     

规模化猪场主要疫病监测及防制

本试验采用酶联免疫吸附试验（ELISA）、正向间接血凝试验（IHA）和乳胶凝集试验（LAT）对曲靖市 16个规模化猪场的2751份血清的8种猪病血清抗体进行了检测。检测结果显示,猪瘟病毒（HCV）、猪O型口蹄疫非结构蛋白（NS）、猪伪狂犬病病毒 gE基因（ADV gE）、猪细小病毒（PPV）、猪日本乙型脑炎病毒（JEV）、猪繁殖与呼吸综合征病毒（PRRSV）、猪圆环病毒2型（PCV 2）和布鲁氏杆菌的血清抗体阳性率分别为0%、0%、22.28%、32.93%、52.99%、29.96%、21.88%和0%;猪瘟（HC）、猪O型口蹄疫（FMD）、猪繁殖与呼吸综合征（PRRS）免疫血清抗体阳性率分别为59.43%、64.56%、48.55%。结果表明,所检测的规模化猪场未感染猪瘟、O型口蹄疫、布鲁氏杆菌病野毒,但存在不同程度地猪伪狂犬病病毒、猪细小病毒、猪日本乙型脑炎病毒、猪圆环病毒2型、猪繁殖与呼吸综合征病毒感染;猪瘟、O型口蹄疫、猪繁殖与呼吸综合征总体免疫效果不佳。     

ELISA和LAT检测猪血清弓形虫抗体的比较

应用弓形虫重组蛋白rMIC3为抗原建立的酶联免疫吸附试验(ELISA)和乳胶凝集试验(LAT)2种方法,平行检测来自36个猪场的471份猪血清中的抗弓形虫特异性抗体。结果显示,ELISA和LAT对母猪弓形虫抗体的阳性检出率分别为42.86%(21/49)和44.90%(22/49),2种方法的阳性检出符合率为95.45%(21/22);对育肥猪的阳性检出率分别为42.06%(135/321)和43.61%(140/321),2种方法的阳性检出符合率为93.57%(131/140);对仔猪的阳性检出率分别为29.07%(30/101)和45.54%(46/101),2种方法的阳性检出符合率为63.04%(29/46)。结果表明,ELISA和LAT可用于猪弓形虫病的诊断和血清学调查,LAT更适合于仔猪弓形虫病的检测。     

Prevalence of Toxoplasma gondii antibodies in stray cats in Sari, northern Iran

Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that can excrete environmentally resistant oocysts. T.gondii is a major zoonotic agent which infects up to one-third of the world population. Toxoplasmosis in neonates and immunocompromised patients can lead to severe disease and death. A cross- sectional parasitological and serological survey with latex agglutination test (LAT) to detect anti-T. gondii antibodies was conducted on 100 serum samples collected from stray cats in five urban areas of Sari, Northern Iran, from April to November 2004. Classification by age, sex, weight, season and region was made. Results analyzed according to specific variables. The overall prevalence of T. gondii IgG antibodies (LAT titre ≥1:1) were found in 40 of 100(40%) of stray cats, with regional variations. Overall 16 of 100(16%) of stray cats had diagnostically significant antibody titres (LAT ≥ 1:64). Prevalence was significantly higher in adult cats (1.5–3.0 kg, 54.5% of 66) than in juvenile cats and kittens (≤1.4 kg, 11.8% of 34) and higher in female stray cats (44.4% of 72) than in male stray cats (28.6% of 28). Toxoplasma seroprevalence was highest in the season of spring (22.4%). There was a significant difference in the prevalence of infection relative to host age and weight (P < 0.05). No significant difference was found between the prevalence of infection relative to host gender, urban sites and season (P > 0.05). Prevalence of T. gondii oocyst was also analyzed by a routine coprological method in 100 cats. T. gondii oocysts were not found in any faecal samples analyzed. Only 2 out of 100 smear preparations of intestinal mucosa showed trophozoites of T. gondii.     

乳胶凝集试验检测鸡传染性法氏囊病毒血清抗体的研究

以鸡传染性法氏囊病病毒(IBDV)VP2基因工程抗原致敏乳胶微粒,用IBDV阳性血清进行方阵滴定,以最佳致敏系件制成乳胶抗源,建立乳胶凝集试验(LAT),用来检测血清中IBD抗体。对467份待测血清分别、同时作LAT和双向琼琼脂免疫扩散试验(DATA)。结果,LAT阳性419份,阴性48份;DAGT阳性425份,阴性42份.试验表明乳胶凝集试验操作简便、快速、敏感性高、特异性强,可用于现场检测,适合基层单位检测IBDV血清抗体。     

Analysis of L-type amino acid transporter in canine hepatocellular carcinoma

Analysis of L-type amino acid transport expression of hepatocellular carcinoma cells (HCCs) of the dog was performed. The leucine transport activity of canine HCCs was 0.628 ± 0.018 nmol/mg protein/min. The inhibitor of LAT 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid (BCH) reduced 90% of the activity at 1 mM. The deduced amino acid sequences of canine LAT2, LAT3 and LAT4 were well conserved in mammalians, exhibiting 89, 88 and 77% homology, respectively. RT-PCR revealed distinct LAT1 expression compared with normal hepatocytes. Western blotting analysis confirmed the potent LAT1 expression in canine HCCs but not hepatocytes, and real-time RT-PCR analysis indicated that canine HCCs possessed 28 times higher LAT1 expression than hepatocytes. These results indicated that the leucine transport activity of canine HCCs was due to LAT1.