旋毛虫肌幼虫囊包染色标本制作方法的改进

目的探讨旋毛虫肌幼虫囊包染色标本的制作方法。方法采用醋酸卡红染色法制作旋毛虫肌幼虫囊包标本。结果醋酸卡红染色的标本,囊包轮廓清晰,囊内幼虫透明度及清晰度高,体态自然,立体感较强。结论醋酸卡红染色可用于旋毛虫肌幼虫囊包永久标本的制作。     

一种旋毛虫肌幼虫囊包标本的单染制作方法

本研究采用醋酸明矾卡红染色液对旋毛虫肌幼虫囊包标本进行染色。为观察不同染色时间对制片效果的影响,分别采用4种染色方法:①染色20 h后,分色;②染色2.5 h,不分色;③染色20 h,不分色;④染色40 h后,分色。结果显示,染色方法 1制作的标本效果最佳。囊包、囊内幼虫和周围肌纤维色泽差别较大,易于观察。囊壁内外两层结构可清晰辨别,囊内幼虫典型。     

旋毛虫肌幼虫囊包不染色标本的制作

在医学寄生虫学实验教学中,制作旋毛虫肌幼虫囊包标本通常采用染色封制法,梭形囊包明显,但如果分色过程掌握不好,虫体轮廓不清。作者尝试制作旋毛虫肌幼虫囊包不染色封制标本,用于教学实践取得良好效果。介绍如下。 旋毛虫肌幼虫囊包取自感染旋毛虫幼虫的猪横纹肌。操作工具:眼科剪,眼科镊,载玻片,大瓶皿,显微镜,吸管,玻片标本盘等。试剂:甲醛,蒸馏水,乙醇,水杨酸甲酯,中性树胶。 将感染旋毛虫幼虫的肌肉剪成碎块,夹在两张载玻片之间,轻轻加压、捆扎,置于10％甲醛固定12～24 h。解开载玻片后再浸泡1～2h,充分固定。倾去甲醛,用蒸馏水洗3次,每     

福寿螺体内广州管圆线虫Ⅲ期幼虫的形态学观察

目的观察福寿螺体内广州管圆线虫Ⅲ期幼虫的形态学特征。方法实验室培养的广州管圆线虫Ⅰ期幼虫感染禁食24h的福寿螺,61d解剖,取其肺囊和足肌,常规制作石蜡切片,观察其Ⅲ期幼虫外部形态和内部结构。结果Ⅲ期幼虫虫体卷曲,头部圆钝,咽管始于头部顶端的口孔,在咽肠连接处与肠管连接,尾部尖,肛管清晰。幼虫皮层为伊红染色,皮层外有一层无色透明的鞘膜。部分虫体尾部出现圆柱体,有些幼虫体内出现亚腹腺、很短的双管子宫等Ⅳ期幼虫早期特征。结论广州管圆线虫Ⅲ期幼虫的形态学特征清晰,对预防广州管圆线虫病流行有一定意义。     

旋毛虫肌幼虫囊包标本的复染制作方法

本研究采用单染和复染两种方法对旋毛虫肌幼虫囊包标本进行染色。单染方法:制片经甲醛、乙醇和冰醋酸溶液固定,用乙醇硼砂卡红染色液(4%硼砂水溶液100 ml,卡红1 g,70%乙醇100 ml)染色。复染方法:制片经甲醛、乙醇和冰醋酸溶液固定,用乙醇硼砂卡红染色液和固绿染色液(固绿0.1 g,95%乙醇100 ml)染色。结果显示,单染标本,囊包与周围肌细胞着色无明显差别,结构不清晰,不易观察,囊内幼虫可辨认。复染标本,囊包结构清晰;囊内幼虫、囊包和肌细胞呈不同的颜色,囊包梭形显著,囊内幼虫特征典型。与单染标本相比,复染标本着色适度,染色效果好,易于观察。     

旋毛虫成虫标本离心管染色方法的改良

过去在制作寄生虫染色标本时，对有些用肉眼难以看到的微小虫体或幼虫，为了能在显微镜下看清楚虫体的内部构造，多采用离心管离心沉淀染色法’‘’进行染色。此方法在染色过程中虫体易大量丢失，且不易掌握虫体的染色、分色、脱水等步骤，最后获得标本染色效果往往不够理想，为寻求好的染色方法，我们参照文献”、‘’，对旋毛虫成虫标本离心管离心沉淀染色法进行了改良，取得了较理想的结果，其方法如下。亚旋毛虫成虫固定在70％酒精中保存。2取一张经酸酒精处理的载玻片，用玻璃棒蘸取一小滴蛋白甘油，滴在载玻片中央，用手指加以涂抹，…     

绦虫永久染色标本的制作技术

目的制作绦虫玻片染色标本用于教学和科研。方法用墨汁染色法制作绦虫妊娠节片玻片标本,卡红染色法制作绦虫成熟节片、头节和囊尾蚴玻片标本。结果妊娠节片、成熟节片、头节和囊尾蚴经染色制片后特征结构明显可见。如妊娠节片染色标本清晰可见染成墨汁颜色的子宫主干和分支状的子宫侧支,节片一侧边缘中部可见突出的生殖腔;成熟节片染色标本均染成红色,内有着色更深的雌雄生殖系统各一套。结论墨汁染色和卡红染色制作带绦虫染色标本效果好,可永久保存。     

3种旋毛虫成虫永久标本染色方法的比较

目的探讨制作旋毛虫成虫染色标本的最佳方法。方法分别使用乙醇硼砂卡红、醋酸卡红、固绿3种染色法制作旋毛虫成虫标本。结果乙醇硼砂卡红染色的标本,颜色鲜艳,结构清晰,另外两种染液染色效果不佳。结论乙醇硼砂卡红染色可用于旋毛虫成虫永久标本的制作。     

华支睾吸虫体内神经递质5-羟色胺的分布

目的 了解华支睾吸虫体内神经递质5--羟色胺(5--HT)的分布规律。方法 从感染麦穗鱼中消化分离华支睾吸虫囊蚴,以灌胃方式感染90只昆明小鼠(40～50个囊蚴/鼠),分别于感染后第10、20、30天安乐死处死小鼠,从肝门静脉收集虫体。醋酸洋红染色后光学显微镜下观察不同发育阶段虫体内各器官发育情况;同时进行5--HT免疫荧光染色,在激光扫描共聚焦荧光显微镜下观察虫体内5--HT的分布情况。结果 醋酸洋红染色后光学显微镜下观察可见,感染后华支睾吸虫d₁₀幼虫中,消化、排泄器官基本发育成熟,生殖器官未完全发育成熟,子宫内可见散在分布的虫卵;d₂₀幼虫中,生殖器官发育成熟,子宫中虫卵数量增多,睾丸分支增多;d₃₀成虫中,消化、排泄、生殖系统均发育成熟,子宫内虫卵较d₂₀幼虫排列更紧密,睾丸增大且分支明显。5--HT免疫荧光染色后激光扫描共聚焦荧光显微镜下可见,d₁₀幼虫的中枢经系统节、神经联合、口吸盘处荧光染色较强,在虫体内部器官中呈点状分布;d₂₀幼虫的口吸盘...     

广州管圆线虫幼虫致小鼠脑组织细胞凋亡研究

目的建立广州管圆线虫幼虫实验感染ICR小鼠动物模型,对其脑组织细胞凋亡情况进行检测。方法对小鼠动物模型脑组织切片,HE染色观察凋亡细胞的形态变化,再利用TUNEL法进行组织细胞凋亡水平检测及Western-blot方法检测不同感染时期小鼠脑组织中Caspase3表达水平变化,对广州管圆线虫幼虫致小鼠脑组织细胞凋亡进行综合评价。结果HE染色发现感染鼠脑组织细胞形态变化,经TUNEL法观察到感染鼠大脑、小脑、脑干部分均有不同程度的细胞凋亡,细胞凋亡率分别为20%、50%和70%,且用Western-blot方法分析Caspase3结果显示随着幼虫感染小鼠时间的延长,活性Caspase3表达量增加,即细胞凋亡程度增加。结论证实广州管圆线虫幼虫可诱导小鼠脑组织细胞凋亡,且随感染时间的延长组织凋亡水平亦增加。     

An assessment of the cost-effectiveness of fast tracking bacteriological specimens for mycobacteria

A virtual model, using six predetermined criteria for fast tracking tuberculosis specimens was devised to improve the cost effectiveness of the MB/BacT system. All specimens received at a central laboratory were audited for the six criteria over a 6-month period. By assuming that only those specimens fulfilling these criteria were fast tracked the theoretical cost savings could be calculated. To prevent possible delay in speciating mycobacteria, the number of criteria were expanded to nine, and a further 6 month audit carried out. In the first 6-month period, 728 specimens were tested. Had the initial hypothetical criteria excluded some of the specimens, only 351 specimens would have been tested through the fast-track system at a saving of pounds sterling 942 (dollars 1696), (52%) of the total cost, but five culture results positive for environmental mycobacteria would have been delayed. In a second 6-month survey the criteria were expanded. Using these no positive culture would have been missed but the savings would only have been 26% of the total cost. Introducing exclusion criteria for rapid testing can improve the cost effectiveness of rapid culture methods with no important loss of clinically necessary information.     

2946份痰标本抗酸杆菌检查结果与咳痰时间的关系

目的　探讨痰标本的咳痰时间对抗酸杆菌检查结果的影响。方法 采用痰直接厚涂片萋-尼氏抗酸染色显微镜(物镜100×)检查,统计不同咳痰时间的痰标本阳性检出率和全部阳性痰标本按咳痰时间的分布。结果 2946份痰标本中,清晨痰、夜间痰、即时痰阳性检出率有显著性差异。462份阳性痰标本中,按清晨痰、夜间痰、即时痰统计分布有非常显著意义。结论 咳痰时间对痰标本抗酸杆菌检出有很大影响,要强调清晨痰的送检。     

PCR快速检测腹泻患者粪中痢疾杆菌致病基因ipaH和ial

目的: 建立一种快速、特异、敏感的分子生物学诊断方法对腹泻患者粪中痢疾杆菌致病基因ipaH及ial进行检测.方法:分别应用常规培养生化血清学鉴定方法、普通PCR、巢氏PCR(nested-PCR)方法, 对71例腹泻患者粪标本进行检测. 痢疾杆菌致病基因ipaH及ial的扩增产物经电泳分离.结果: 在71例腹泻患者粪标本中, 常规培养生化血清学鉴定方法分离出福氏痢疾杆菌24例, 宋内痢疾杆菌23例, 志贺痢疾杆菌1例, 沙门菌23例. 采用普通PCR法检测痢疾杆菌粪标本48例, ipaH基因均阳性(100%);ial基因阳性34例(70.8%), 余14例为阴性. 对该14例粪标本进而采用巢氏PCR法进行ial基因扩增, 其中12份标本阳性. 48例痢疾杆菌粪标本中46例ipaH和ial基因为双阳性(46/48). 沙门菌粪标本23例中ipaH及ial基因表达均为阴性. 以上两种方法的诊断一致性检验Kappa = 0.937, 差异有统计学意义(P<0.05). 71例腹泻患者粪标本采用PCR和巢氏PCR法扩增得到的ipaH和ial基因特异片段与基因库中发表的标准菌株序列比较, 一致性为100%.结论:建立了快速诊断腹泻患者粪中痢疾杆菌致病基因ipaH和ial的PCR检测方法, 具有简便、快速、特异和敏感的特点.     

山东内陆人体隐孢子虫病调查报告

用改良抗酸染色法检查人群粪便标本，对病人进行治疗随访和流行病学调查。对山东内陆人体隐孢子虫感染情况进行调查防治研究，共检查人群粪便标本3631份，阳性36例，平均人群感染率为1．0％，其中成人为0．2％，5岁及以下婴幼儿为1．2％。婴幼儿是高发年龄组，而且有明显的季节性，男、女性别间无明显差异，以鲁南较多，散发，有村庄聚集性。腹泻为稀糊样便是本病的主要临床表现。结果显示，改良抗酸染色法是比较适用的诊断方法，卵囊的多少能评价腹泻的轻重，隐孢子虫感染多见于腹泻的婴幼儿。作还对流行因素、临床表现、诊断和治疗进行了探讨。     

The reliability of acid fast stained smears of gastric aspirate specimens

A 23-year retrospective study of acid fast stains of unconcentrated gastric aspirate specimens revealed a specificity of 99% and a sensitivity of 30%. All positive smears were from patients with culture-proven mycobacterial disease. Although the sensitivity is low, a positive smear allows a significantly earlier identification of patients with mycobacterial infections when compared to culture.     

The value of fluorescence microscopy of auramine stained sputum smears for the diagnosis of pulmonary tuberculosis

Laboratory diagnosis of pulmonary tuberculosis rests on the bacteriological examination of sputum smears stained by the Ziehl-Neelsen (ZN) method for acid fast bacilli (AFB). In the present study, we have compared light microscopy of ZN stained smears with that of fluorescence microscopy of sputum smears stained by auramine-phenol flurochrome dye for detection of AFB in sputum specimens. Sputum specimens from a total of 2,600 clinically suspected and diagnosed cases of pulmonary tuberculosis were examined by both the methods. Sputum specimens from a total of 1,104 patients were found to be positive for AFB. These included sputa from 975 (37.5%) patients positive for AFB by both ZN and auramine staining methods and sputa from an additional 129 (4.96%) patients positive for AFB by auramine staining only. Thus auramine staining of sputum smears in comparison to that of ZN staining is a better method of sputum microscopy for demonstration of AFB in sputum specimens. Fluorescence microscopy is relatively more sensitive and has the added advantage of allowing a large number of sputum specimens to be examined in a given time, in laboratories equipped with a fluorescent microscope.     

"Nose-blow" smears in multibacillary leprosy patients

332 nose-blow specimens have been examined from 73 untreated multibacillary patients of leprosy before and periodically after they were put on a maximal, minimal or an intermediate multi-drug regimen. 80% of these specimens were found to be positive for acid fast bacilli initially. Bacillary positivity rate was more in samples containing pus or blood. Bacilli were seen in LL, LI as well as BL patients. Nearly half of the cases became negative for AFB in their nose-blow specimens within 3 months of initiation of treatment whereas none of these patients has become negative in skin smears. However, a few cases have continued to discharge bacilli in their nasal secretions even after 12 months of multi-drug regimen therapy.     

Acid fast staining versus ELISA for detection of Cryptosporidium in stool

A prospective study was undertaken to compare the efficacy of ELISA in detection of Cryptosporidium Specific Antigens (CSA) in stool specimens of patients attending various OPDs in a Delhi Hospital. A total of 216 consecutive faecal specimens were examined microscopically after modified acid fast staining. An ELISA was also performed using ELI-WFLL CRYPTO detection kit following the manufacturer's instructions for detection of CSA in stool specimens. Taking microscopy as the gold standard ELISA was found to be 100% sensitive and 99.07% specific in detection of Cryptosporidium spp. The test is easy to perform and interpret.