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首页> 外文期刊>The Internet Journal of Tropical Medicine >Evaluation Of Microscopic Staining Techniques For The Diagnosis Of Opportunistic Protozoan Infections In A Developing Country
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Evaluation Of Microscopic Staining Techniques For The Diagnosis Of Opportunistic Protozoan Infections In A Developing Country

机译：显微染色技术在发展中国家机会性原生动物感染诊断中的评价

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Trichome staining method was superior to other techniques in detecting opportunistic protozoan infections: 35.7% for Cryptosporidium sp, and 50% for Microsporidium sp. The Acid Fast (Carbol Fuchsin) stain was useful only in detecting the oocysts of Cryptosporidium sp.. With 35.7% positivity. Laboratories in developing countries can put Tricrome stain in their priority purchase. Introduction The prevalence of intestinal parasites has been studied extensively in both rural and urban areas of Nigeria 1,2,3. Routine diagnosis of intestinal parasitic infections is usually performed by a simple smear in normal saline or iodine stain solution using light microscopy4. More recently, some unusual protozoans such as Cryptosporidum Spp, Isospora belli, Microspordial Spp, Cyclospora cyatenensis and Blastocystis hominis have become important aetiological agents of diarrhea in children and patients with some type of immune compromised conditions e.g. HIV/AIDS5. In these immunocompromised patients, auto-limiting diarrhea in immunocompetent persons may cause profuse diarrhea, malabsorption syndrome, anorexia, weight loss; low grade fever and abdominal pains or cramps.6,7. With these new frequent presentations, a new pattern of diagnosis of intestinal infections is therefore required. However, in most laboratories, the diagnosis of these unusual protozoan is not part of the routine wet smear ova and parasite light microscopy, but involves the use of special stains.8 The none-use of these stains usually present a challenge in the diagnosis of these uncommon parasites in most developing countries. Therefore, it became necessary to evaluate common microscopic staining techniques available in our laboratory, with the view of recommending a stain or combination of stains that has a broad range application for the various parasites. The Study 70 diarrhoeal stool samples (each pooled from three consecutive days) obtained from various patients, including HIV/AIDS patients who were attending various clinics in Lagos, Nigeria were processed for routine microscopic examinations of ova and parasites. Saline and Iodine wet mount, and permanent slides of Giemsa, Carbol Fuchsin (Kinyoun-modified acid fast) and Weber's modified Trichrome stains were also made. Faecal smears on slides and formal-ether concentration methods were prepared for each stool samples using standard procedures 9,10. The micrometer was used for the accurate measurement of oocysts, cysts, and ova that were seen. The 100X oil immersion lens was employed in all cases to demonstrate the morphology of oocysts. Informed consent was obtained from selected patients after explaining the aim of the study. These patients were already enrolled in an on-going HIV/AIDS clinics and treatment programme.Intestinal opportunistic protozoans, particularly Coccidian and Microsporidum spp. are emerging infections that are important complications of intestinal morbidity in immune compromised patients 10. The laboratory diagnosis of these parasites in our region involves mainly light microscopy, and therefore places a high responsibility on the laboratory scientist. The use of light microscopic stains for diagnosis of these infections has improved greatly over the years, following the modification of some basic staining techniques. This is of importance to laboratories in developing communities, were lack of electron microscopy and molecular biology facilities for the diagnosis of opportunistic infections are lacking. However, the problems of the unavailability of the stains and the cost of purchasing or ordering large varieties are issues facing laboratories in most developing countries.
机译：Trichome染色法在检测机会性原生动物感染方面优于其他技术：隐孢子虫占35.7％，微孢子菌占50％。耐酸性（Carfu Fuchsin）染色仅用于检测隐孢子虫的卵囊。阳性率为35.7％。发展中国家的实验室可以优先购买Tricrome染色剂。引言在尼日利亚1,2,3的农村和城市地区，对肠道寄生虫的流行进行了广泛的研究。肠道寄生虫感染的常规诊断通常是通过使用光学显微镜在生理盐水或碘染色液中简单涂片进行的。最近，一些不寻常的原生动物，如隐孢子虫，贝氏孢子，微孢子虫，Cyatenensis和人乳杆菌已成为儿童和某些类型的免疫功能低下的患者腹泻的重要病原体。艾滋病毒/艾滋病5。在这些免疫功能低下的患者中，免疫能力强的人的自限性腹泻可能引起大量腹泻，吸收不良综合征，厌食，体重减轻;低烧和腹痛或痉挛6.，7。因此，随着这些新的频繁出现，需要一种新的诊断肠道感染的模式。但是，在大多数实验室中，这些异常原生动物的诊断不是常规湿涂片卵和寄生虫光学显微镜检查的一部分，而是涉及使用特殊的染色剂。8不使用这些染色剂通常在诊断血吸虫病方面存在挑战。在大多数发展中国家，这些罕见的寄生虫。因此，有必要评估我们实验室中可用的常规显微染色技术，以期推荐一种对各种寄生虫具有广泛应用的染色剂或染色剂组合。研究对70例腹泻大便样本（每个样本连续三天收集），这些样本取自各种患者，包括在尼日利亚拉各斯各诊所就诊的HIV / AIDS患者，进行了常规的卵和寄生虫显微镜检查。还制作了盐水和碘湿法固定件以及Giemsa，Carbol Fuchsin（Kinyoun修饰的耐酸碱）和Weber修饰的Trichrome染色剂的永久载玻片。使用标准程序9,10为每个粪便样品准备载玻片上的粪便涂片和形式醚浓缩方法。千分尺用于准确测量观察到的卵囊，囊肿和卵子。在所有情况下均使用100X油浸镜头来证明卵囊的形态。在解释了研究目的之后，从选定的患者中获得了知情同意。这些患者已经参加了正在进行的艾滋病毒/艾滋病诊所和治疗计划。肠道机会性原生动物，特别是球虫和小孢子菌。这些是免疫受损患者肠道疾病的重要并发症，即新兴感染。10在我们地区，这些寄生虫的实验室诊断主要涉及光学显微镜，因此对实验室科学家负有重大责任。多年来，随着一些基本染色技术的改进，使用光学显微镜染色剂诊断这些感染的方法已大大改善。这对发展中社区的实验室很重要，因为缺少电子显微镜和分子生物学设施来诊断机会性感染。但是，污渍不可用以及购买或订购大品种的成本问题是大多数发展中国家实验室面临的问题。

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