• Parasites, Hosts and Diseases
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• 제47권4호
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• pp.353-357
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• 2009

Cryptosporidium can cause gastrointestinal diseases worldwide, consequently posing public health problems and economic burden. Effective techniques for detecting contaminated oocysts in water are important to prevent and control the contamination. Immunomagnetic separation (IMS) method has been widely employed recently due to its efficiency, but, it is costly. Sucrose floatation technique is generally used for separating organisms by using their different specific gravity. It is effective and cheap but time consuming as well as requiring highly skilled personnel. Water turbidity and parasite load in water sample are additional factors affecting to the recovery rate of those 2 methods. We compared the efficiency of IMS and sucrose floatation methods to recover the spiked Cryptosporidium oocysts in various turbidity water samples. Cryptosporidium oocysts concentration at 1, $10^1$, $10^2$, and $10^3$ per $10{\mu}l$ were spiked into 3 sets of 10 ml-water turbidity (5, 50, and 500 NTU). The recovery rate of the 2 methods was not different. Oocyst load at the concentration < $10^2$ per 10 ml yielded unreliable results. Water turbidity at 500 NTU decreased the recovery rate of both techniques. The combination of sucrose floatation and immunofluorescense assay techniques (SF-FA) showed higher recovery rate than IMS and immunofluorescense assay (IMS-FA). We used this SF-FA to detect Cryptosporidium and Giardia from the river water samples and found 9 and 19 out of 30 (30% and 63.3%) positive, respectively. Our results favored sucrose floatation technique enhanced with immunofluorescense assay for detecting contaminated protozoa in water samples in general laboratories and in the real practical setting.

• Parasites, Hosts and Diseases
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• 제23권2호
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• pp.285-292
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• 1985

The present study has been designed as a basic study on laboratory diagnosis of giardiasis and to demonstrate a more effective method for the detection of Giardia lamblia cyst with the inherent advantages of minimizing both the number of stool examinations required and the interval of stool collections for estimating the real state of prevalence in the shortest time possible. There were 3 subject groups of 75 children each currently residing in an orphanage in Gunsan city, Jeonbuk province from which stool specimens were collected every day, every other day, and every 3 days. The procedure is as follows: 1) resuspend the InKed sample after fixation with SAF solution 2) centrifuge the sediment for 1 min. at 2, 000 rpm after straining through gauze into a tube 3) divide the sediment into 3 parts and use them for direct fecal smear, formalin-ether concentration (MGL) and zinc sulfate ($ZnSO_4$) floatation techniques. The results are summarized as follows: 1. Overall infection rate after 10 trials showed a 60% positive indication. The positive rate among children under 4 years old was significantly higher than the rate in children over 4 years old. No significant difference in rate by sex was observed. 2. The results of eBaminations by direct fecal smear and MGL techniques appeared more accurate than that obtained by $ZnSO_4$ ftoatation method as indicated by a higher positive rate. Of all three methods concerned, combinations of two demonstrated a higher positive rate than that shown by any one alone. 3. In three consecutive examinations under varying conditions such as different days, the cyst detection rate by MGL technique indicated 83%. In 5 examinations under the same varying conditions, the indicated rate was 94%. 4. The interval of stool collection proved to be insignificant for the cyst detection rate. In conclusion, both MGL method and modified fecal direct smear can provide a good cyst detection rate of G. lamblia provided that more than 3 consecutive examinations of stool under varying conditions are carried out.

• 농촌의학ㆍ지역보건
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• 제4권1호
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• pp.62-70
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• 1979

In order to evaluate the present status of hook-worm infection in the rural areas in Korea, stool examinations were undertaken in several selected rural villages in each year of 1970 and 1977. The methods employed were cellophane thick smear technique and brine floatation technique in 3,454 specimens collected from 7 different rural villages for the prevalence rate of hookworm infection and Stoll's dilution egg counting technique for the intensity of hookworm infection. The incidences of Ancylostoma duodenale and Necator americanus infections were determind by polyethylene tube coproculture and by the evacuated adult worms after treatment with anthelmintics to the patients who were infected by hookworms. The results are as follows : 1) In 1970, 323(18.6%) out of 1,737 specimens collected from 4 different rural areas were positive for hook worm infection and the ranges of prevalence rates in those suveyed areas were shown as 6.9 to 38.2 per cent. On the other hand, 144(8.2%) out of 1,717 specimens collected from 3 different rural areas in 1977 were shown as positive for hookworm infection and 7.5 to 10.6 per cent of prvalence rates were shown in the surveyed areas. 2) In the age group under 10 years, there was no infection of hookworm in the studied areas in 1977, however in 1970 the prevalence of the hookworm infection was shown as very low in the age group of 5~9. On the whole, the hookworm prevalence steadily increased from the age of 15 in the studied areas in 1970, however in 1977 the prevalence increased from the age of 20 to the older age groups. 3) There was not observed much differences in the intensity of hookworm infection among the studied areas in 1970 and 1977. The mean EPG in each studied areas were shown under 1,000 EPG and mean numbers of evacuated worms per infected persons were counted as 9.4 to 15. 1. 4) Both A.duodenale and N.americanus have been found in all studied areas but the incidences and st-ate of distribution of two species of hookworn were shown as different in each localities.

• 생명과학회지
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• 제19권6호
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• pp.728-734
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• 2009

본 연구에서는, PBMC로부터 고순도 및 고수율의 단세포를 분리하기 위해 Histopaque 용액의 density 및 osmolarity를 PBS와 NaCI 용액으로 1.072 g/ml 및 335 mOsm로 조절하여 간단하고 경제적인 방법인 부유밀도구배 원심분리법을 개발하였다. 위와 같이 조절된 Histopaue 용액을 사용하여 부유밀도구배 원심분리를 시행한 결과 단세포는 Top층에서 가장 많이 회수되었다. 회수된 단세포의 순도는 71.44${\sim}$82.38%의 범위를 나타내었고 평균 74.75${\pm}$3.84% 이었다. 또한, 수율은 21.02${\sim}$53.63%의 범위를 나타내어 평균 32.62${\pm}$11.16% 이었다. 이러한 방법의 부유밀도구배 원심분리에 의해 분리된 단세포를 수지상세포로 분화 유도시켜 그 형태, 표현형, 기능적인 면을 분석해 보았을 때 전형적인 수지상세포의 기능을 나타내었다. 결론적으로, PBMC로부터 단세포를 분리하기 위해 본 연구에서 사용된 부유밀도구배 원심분리 방법은 plastic adherence, elutriation 및 immunomagnetic selection보다 용이하고 경제적인 방법이다. 나아가 이러한 장점을 바탕으로 임상단계에서 수지상세포를 안전하고, 효율적으로 배양할 수 있는 closed system에도 적용이 가능할 것으로 기대된다.