4种狂犬病病毒ELISA抗体检测试剂盒的评价

为评价国产狂犬病病毒ELISA抗体检测试剂盒的性能,选择4种国产品牌ELISA抗体检测试剂盒,分别检测463份已用荧光抗体病毒中和试验（FAVN）测知狂犬病病毒抗体效价的犬血清,用Kappa检验和配对卡方检验评价ELISA试剂盒与FAVN检测结果的一致性和差异性,计算ELISA试剂盒的诊断敏感性、诊断特异性、符合率和重复性等,并比较4种试剂盒的使用范围、样品稀释倍数等参数。结果显示：经Kappa检验,4种试剂盒与FAVN一致性均为弱;经配对卡方检验,A、B试剂盒与FAVN检测结果的差异不显著（P> 0.05）,而C、D试剂盒差异显著（P <0.05）。诊断敏感性,D试剂盒最高,B试剂盒最低;4种试剂盒诊断特异性均较低,为29.5%～57.7%;各试剂盒符合率相当,为71.3%～72.8%。综合敏感性、特异性、重复性等考量因素,得出C试剂盒更能满足免疫后抗体检测需求,A试剂盒也较好,两者可作为候选试剂盒。结果表明,4种国产狂犬病病毒ELISA试剂盒的诊断敏感性、诊断特异性、批间稳定性等性能需进一步提高,以满足基层免疫抗体监测需求。     

禽巴氏杆菌B_(26)-T_(1200)弱毒冻干苗的研究 Ⅰ.禽巴氏杆菌B_(26)- T_(1200)弱毒菌株的选育

从广西各地患禽巴氏杆菌病急性死亡的鸡、鸭肝脏分离出５６株禽巴氏杆菌强毒株,经培养特性和生化特性试验鉴定,从中选出１５株典型禽巴氏杆菌菌株。Ｃａｒｔｅｒ荚膜抗原分型鉴定,１５株菌均为荚膜Ａ型,间接血凝滴度为１∶１６０～１∶６４０。通过毒力和免疫原性测定,从中选择毒力较强、免疫原性较好的Ｂ２５、Ｂ２６、Ｂ２７３菌株进行人工致弱,其中Ｂ２６菌株在０．１％裂解全血马丁汤中,通过物理诱变方法致弱,即在传代过程中,把培养温度由３７℃逐渐提高到４５℃,每１２ｈ传１代而成功致弱,传至１２００代时,毒力显著减弱,且仍保持其良好的免疫原性和培养特性,从而获得了禽巴氏杆菌Ｂ２６－Ｔ１２００弱毒菌株     

丝胶凝胶物理性状的研究

丝胶在凝胶化时,强力增加,而表面张力减小。丝胶凝胶,在30～40℃热水处理时,溶解量很小;50一70℃热水处理时,溶解量增多,溶解速度呈恒定状态;80℃以上,呈快速溶解。根据丝胶凝胶的DTA分析,其吸热分解峰向高温侧移动,这表明丝胶分子的有序性、结晶度提高了。丝胶凝胶在热水处理时,转化为液胶,同时丝胶分子也由β构象转化为无视卷曲结构。并且,丝胶液胶又可重新凝胶化。     

大豆卵磷脂分离纯化工艺的研究进展

大豆卵磷脂是以大豆油脚为原料提炼的卵磷脂,含有胆碱、维生素、矿物质、亚麻酸及次亚麻酸等成分,具有多种生理功能,并在许多领域内得到了广泛的应用。然而由于技术原因,高纯度卵磷脂的生产仍存在一定的技术障碍,阻碍了其在医药等领域的推广应用。为此,对近年来大豆卵磷脂分离纯化工艺的研究进展做一简要综述和分析,以期为高纯度大豆卵磷脂的研究与应用提供参考。     

黄皮茎段初代培养物的建立

表面污染是木本植物组织培养污染率高的主要原因。选择晴、阴天气下生长的试材或者采用茎尖培养,是降低污染率、成功建立无菌初代培养物的有效措施。适合黄皮茎段初代培养的条件为:WPM+BA1.0+KT0.5+NAA0.5+GA_30.25+4～5％蔗糖。     

Expression in Escherichia coli of open reading frame gene segments of HTLV-III

Human T-cell lymphotropic virus type III (HTLV-III), the causative agent of the acquired immune deficiency syndrome (AIDS), was recently isolated and its genomic structure analyzed by DNA cloning methods. In the studies reported here a combined cloning and expression system was used to identify HTLV-III encoded peptides that react immunologically with antibodies in sera from AIDS patients. Cloned HTLV-III DNA was sheared into approximately 500-base-pair fragments and inserted into an "open reading frame" expression vector, pMR100. The inserted DNA was expressed in Escherichia coli transformants as a polypeptide fused to the lambda CI protein at its amino terminus and to beta-galactosidase at its carboxyl terminus. Sera from AIDS patients containing antibodies to HTLV-III were then used to screen for immunoreactive fusion proteins. Twenty clones, each specifying a fusion protein strongly reactive with AIDS serum, were identified. DNA sequence analysis indicated that the HTLV-III fragments were derived from the open reading frame DNA segments corresponding to the gag and pol gene coding regions and also the large open reading frame region (env-lor) located near the 3' end of the viral genome.     

稻-鸭生态系统中二化螟消长动态研究

2003年4～10月,在稻鸭共栖田中,对二化螟发生规律的系统研究结果表明,稻田养鸭能显著减轻二化螟的为害。与对照相比,中稻田养鸭区二化螟为害株率减少13.4%～47.1%;晚稻田减少62.2%。稻鸭共栖能减少化学农药的使用量,具有较好的经济、社会和生态效益。     

Three novel single-nucleotide polymorphisms of MBL1 gene in Chinese native cattle and their associations with milk performance traits

Mannan-binding lectin (MBL), a pattern recognizing serum protein, participates in the innate immune system of mammals as an opsonin. In humans, single-nucleotide polymorphisms (SNPs) in MBL2 gene were found to cause various innate immune dysfunctions. In the present study, we discovered three single-nucleotide polymorphisms of the MBL1 gene in Chinese native cattle and analyzed their associations with milk traits. By screening the genetic variation of MBL1 in 1053 individuals of three Chinese native cattle breeds including China Holstein, Luxi Yellow and Bohai Black using created restriction site–polymerase chain reaction (CRS–PCR), PCR–restriction fragment length polymorphism (PCR–RFLP) and DNA sequencing techniques, three new SNPs, g.855G>A, g.2651G>A and g.2686T>C, were found to have allele frequencies of 0–12.65%, 24.07–42.39% and 56.95–73.68%, respectively. While SNP g.855G>A is located within intron ?, the other two SNPs reside in the exon II region with one mutation being non-synonymous (GTT (Val) > ATT (Ile)) and the other synonymous (GCT (Ala) > GCC (Ala)). Among the 596 Chinese Holstein cattle with at least 3 lactation Dairy Herd Improvement (DHI) records, eight different haplotypes and 19 genotype combinations were detected. Statistical analyses revealed no correlation between either g.855G>A or g.2686T>C and somatic cell score (SCS), however significant association was found between g.2651G>A and SCS, suggesting a possible role of this SNP in the host response against mastitis. Our data also suggested that the combined genotypes of GGC/AAC with the lowest SCS, AAT/AAT with the highest protein content and AGC/AGC with the highest 305-d milk yield were favorable combinations for mastitis resistance and milk production traits. Therefore, GGC/AAC, AAT/AAT and AGC/AGC can be used as possible candidates for marker-assisted selection in the dairy cattle breeding program.     

不同单酶制剂混合检测对比研究

试验以木聚糖酶、纤维素酶、淀粉酶和酸性蛋白酶为对象,首先研究了酸性蛋白酶分别和其他3种单酶进行混合后酶活测定值变化情况,结果表明,酸性蛋白酶对其他3种单酶的检测结果没有影响;试验第二部分研究了4种单酶整体混合在一起后各单酶测定值的变化情况,结果表明,木聚糖酶测定值出现很大偏高,其他3种单酶测定结果未出现显著偏差;进行进一步论证试验表明,淀粉酶可能是使木聚糖酶测定结果偏高的原因。     

美国车厘子组织培养试验

采用美国思加图车厘子室内盆栽萌发的嫩芽和老芽作为外植体,分析不同处理时间对车厘子外植体萌芽效果的影响;采用不同浓度的植物生长调节剂对其进行诱导增殖和生根培养,筛选最佳组培方法。结果表明：嫩芽是车厘子外植体的最佳选择,通过7 s酒精和0.1%升汞9 min消毒获得无菌体,污染率低、褐化率较低、萌芽率60%。芽诱导培养基为MS+6-BA 1.0 mg/L+NAA 0.2 mg/L,增殖系数为3.92。生根培养基为1/2MS+NAA0.6 mg/L,生根率为100%。移栽炼苗基质比例为V（腐殖土）∶V（红土）=4∶6,盖膜驯化,保湿温度（25±2）℃,湿度80%,成活率达90%以上。