• Parasites, Hosts and Diseases
• /
• 제56권2호
• /
• pp.113-119
• /
• 2018

Cryptosporidium species is an important cause of gastrointestinal infections globally. This study aimed to shed light on its role in diarrheic immunocompetent patients in Beni-Suef, Egypt and to compare three diagnostic methods. Two hundred diarrheic patients, $37{\pm}16.8year$ old, were enrolled. Stool samples were examined by light microscopy, using modified Ziehl-Neelsen stain (MZN) for Cryptosporidium spp. oocysts. Coproantigens were detected by sandwich ELISA. DNA molecular diagnosis was done by nested PCR. PCR yielded the highest detection rates (21.0%), compared to ELISA (12.5%) and MZN staining method (9.5%). The higher infection rates were in 20-40 year-old group, followed by 40-60 year-old. Association between epidemiologic factors was statistically not significant; positivity and gender, clinical manifestations, residence, source or water, or contact with animals. Cryptosporidiosis is an important enteric parasitic infection in Beni-Suef and PCR remains the gold standard for diagnosis.

• 대한의용생체공학회:의공학회지
• /
• 제23권3호
• /
• pp.235-241
• /
• 2002

결핵을 진단하는 방법 중에서 신속하고 비교적 비용이 적게드는 방법은 객담을 통한 결핵균 도말 검사이다. 결핵균 도말 검사는 슬라이드에 도말한 환자의 객담을 가온 과정을 통해 고착시키고. acid-fast 염색방법을 통해 염색시킨 후 현미경으로 결핵균을 관찰하는 것이다. Acid-fast 염색방법은 크게 hot staining과 cold staining 방법 두 가지가 있으며, 우리나라에서는 염색 결과가 선명한 hot staining 방법인 Ziehl-Neelsen 방법을 주로 이용한다. 그러나, 기존의 결핵균 자동염색기는 가온 기능이 없어 환자의 객담을 슬라이드에 검사자가 고착을 시켜야 하고. 선명도도 낮은 문제점을 가지고 있다. 본 연구에서는 검사자의 인력 절감과 검사자 개인의 염색 능력에 따른 염색 정도의 변화를 줄이기 위해 가온이 가능한 결핵균 자동염색기를 개발하였다 개발된 염색기는 객담의 고착에서 염색 그리고 건조가지 전 과정이 자동으로 이루어진다. 염색 시간은 5개의 슬라이드를 고품질로 염색할 경우 21분이 소요되었다. 성능 평가를 위해 총 91개 객담을 대상으로 자동과 수동염색을 시행하여 일치율을 비교해 본 결과 75%로 통계적으로 유의한 차이를 보이지 않았다 (P>0.05).

• 대한의생명과학회지
• /
• 제23권2호
• /
• pp.96-103
• /
• 2017

Nocardia species (spp.) are opportunistic pathogen in immunocompromised hosts. The genus Nocardia contains more than 70 species. Nocardia takedensis has been recently reported as a new species of the genus Nocardia. In this study, we describes the first clinical isolate of N. takedensis from the skin lesion in Busan, Korea. For the identification of clinical isolate to the species level as N. takedensis, classical methods (colony morphology, biochemical characteristics, and antimicrobial susceptibility), molecular method (16S rRNA gene sequencing), and MS (mass spectrometry) analysis were conducted. Clinical isolates grew slowly on the culture media (5% sheep blood agar and chocolate agar) under 5% $CO_2$ condition. Especially, carotene pigmentation was detected well on the media. Using mass spectrometry, Nocardia isolate was not identified to the species level. However, molecular method based on 16S rRNA sequencing confirmed the isolate as N. takedensis correctly. N. takedensis isolate was partial positive for acid-fast bacilli on the Ziehl-Neelsen method. And it was observed to be resistance to amoxicillin/clavulanic acid and ciprofloxacin. Our results provide useful information to develop optimal identification protocol of N. takedensis in clinical diagnostic laboratories.

• Parasites, Hosts and Diseases
• /
• 제50권1호
• /
• pp.57-62
• /
• 2012

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children ($P$<0.0001). The most common protozoan infection in immunosuppressed group was $Cryptosporidium$ $parvum$ (60.2%), followed by $Blastocystis$ $hominis$ (12.1%), $Isospora$ $belli$ (9.7%), and $Cyclospora$ $caytenensis$ (7.8%). On the other hand, $Entamoeba$ $histolytica$ (24.6%) and $Giardia$ $lamblia$ (17.6%) were more common than other protozoans in immunocompetent children.

• 대한수의학회지
• /
• 제47권4호
• /
• pp.443-447
• /
• 2007

A 7-year-old, female snow goose (Anser caerulescens hyperboreus) with history of decreased activity for 2 month died in Daejeon Zoo Land in September 2006. At necropsy, granulomatous pneumonia and hepatomegaly with multiple cysts were observed. Small masses were found in the spleen. Microscopically, fibrinous pneumonia distributed in most of the lung lobe with pulmonary edema and congestion. Especially, granulomatous inflammation with numerous multinucleated giant cells was observed around the dilated bronchi. To confirm the diagnosis, acid-fast (Ziehl-Neelsen method) and periodic acid-Schiff (PAS) staining was performed. Acid-fast staining showed red bacterial colony indicating tuberculosis. PAS staining was also positive enough to diagnose aspergillus spp. co-infection that was an opportunistic fungi occurring in immuno-compromised animals. Based on the above results, we confirmed that the case submitted was diagnosed as avian tuberculosis.

• 대한의생명과학회지
• /
• 제22권1호
• /
• pp.24-28
• /
• 2016

Tuberculosis (TB) remains an unsolved community health problem since identification of its causing microorganism called Mycobacterium tuberculosis (MTB) by Robert Koch in 1882. Annually, eight million TB cases are newly reported and 2~3 million patients die from TB. Pulmonary TB is highly infectious and untreated pulmonary TB patients are believed to infect >10 people in a year. The conventional methods for diagnosis of TB are chest X-ray and isolation of the causing microorganisms from patient specimens. Screening of TB is conducted with smeared sputum in slides, and TB is confirmed by identification of MTB in cultured specimens. One of the fatal pitfalls of screening detection for smeared sputum is that it is impossible to distinguish MTB and other acid-fast bacilli (AFB) because they are stained equally with Ziehl-Neelsen (ZN) stain. Culture of MTB is the most reliable method for diagnosis of TB but it takes 4~8 weeks. In this report, we suggest a fast and highly-reliable MTB detection method that distinguishes AFB in sputum samples. Purified DNA from the AFB stained slide samples offered by The Korean Institute of Tuberculosis were used to detect infected MTB in patients. PCR, real-time PCR and reverse blot hybridization assay (REBA) methods were applied to purified DNA. Conclusively, the real-time PCR method was confirmed to produce high sensitivity and we were able to further detect drug-resistant MTB with REBA.

• 대한의생명과학회지
• /
• 제19권2호
• /
• pp.153-157
• /
• 2013

Although culture is the gold standard method to identify mycobacteria, its use in tuberculous lymphadenitis (TBL) is limited due to formalin fixation of the submitted specimens. We evaluated the performance of quantitative real-time PCR (q-PCR) for Mycobacterium Tuberculosis (MTB) in granulomatous lymphadenitis using formalin-fixed paraffin-embedded (FFPE) tissues. From 2000 to 2010, a total number of 117 cases of lymph node samples with granulomatous inflammation which were surgically removed and fixed in formalin were studied. Hematoxylin & Eosin (H&E) and Ziehl-Neelsen-stained (ZN) slides were reviewed. qPCR using Real TB-Taq$^{(R)}$ was performed for all cases to identify Mycobacterium tuberculosis. Thirteen non-tuberculous lymphadenopathy cases were used as negative control. Cervical lymph nodes were more frequently affected (60%, 70/117) than other sites. ZN stain for acid fast bacilli was positive in 19 (16.24%) cases. qPCR for tuberculosis was positive in 92 (78.63%) cases. Caseous necrosis was found in 103 (88.03%) cases. While the ZN stain and qPCR were both negative in all control cases, the qPCR showed a significantly higher positive rate (78.63% vs. 16.24%) compared to ZN stain in histologically diagnosed TBL. Quantitative real-time PCR proves to be more sensitive than ZN stain for diagnosis of tuberculous lymphadenitis.

• 대한두경부종양학회지
• /
• 제12권2호
• /
• pp.169-176
• /
• 1996

결핵의 진단은 특징적 병리조직양상, 항산균 염색에 의한 균 증명과 M. tuberculosis균 배양으로 이루어지나 형태적으로는 결핵이 의심되더라도 항산균이 조직표본이나 도말에서 검출되지 않거나 M. tuberculosis가 배양되지 않아 정확한 원인적 진단이 불가능할 경우가 많다. 이에 저자들은 경부 결핵성 임파선염으로 적출되어 보내어진 경부 임파선 조직의 신선한 조직이나 통상적으로 처리되어 제작된 파라핀 블록을 이용하여 M. tuberculosis에 특수한 반복성 DNA sequence인 IS986를 표적으로 한 primers을 사용하여 nested PCR방법을 이용하여 예민도가 높은 M. tuberculosis 검출로 빠른 시간 내에 결핵을 진단하고자 본 연구를 실시하였다. 최근 유전자 기술의 진보로 M. tuberculosis의 여러 항원들의 유전자가 클론화되고 그 염기 배열이 밝혀졌으며 이에 저자들은 결핵의 확진을 위하여 파라핀 포매조직을 대상으로 nested PCR에 의한 188bp의 DNA를 증폭한후 증폭한 DNA분절의 염기 배열을 결정한 후 Bst UI와 Hha I 효소를 이용한 소화과정을 거친 후 restriction fragment length polymorphism(RFLP)을 은염색에 의해 그 패턴에 의해 M. tuberculosis를 확인하고 또한 다른 종의 Mycobacteria를 배제시킬 수 있었다. 본 방법은 1$\sim$2일에 끝나며, 방사선물질을 사용하지 않으면서도 감도 및 특이성이 우수하여 일반 병리실힘실에서도 M. tuberculosis를 포함한 각종 항산균의 신속한 검출법으로 손쉽게 사용할 수 있다고 생각되어 이에 보고하였다.

• Tuberculosis and Respiratory Diseases
• /
• 제79권4호
• /
• pp.282-288
• /
• 2016

Background: Tuberculosis (TB) is a major health problem, and accurate and rapid diagnosis of multidrug-resistant (MDR) and extended drug-resistant (XDR) TB is important for appropriate treatment. In this study, performances of solid and liquid culture methods were compared with respect to MDR- and XDR-TB isolate recovery and drug susceptibility testing. Methods: Sputum specimens from 304 patients were stained with Ziehl-Neelsen method. Mycobacterium tuberculosis (Mtb) isolates were tested for recovery on $L{\ddot{o}wenstein$-Jensen (LJ) medium and the BacT Alert 3D system. For drug susceptibility testing of Mtb, isolates were evaluated on M-KIT plates and the BacT Alert 3D system. Results: The recovery rates were 94.9% (206/217) and 98.2% (213/217) for LJ medium and the BacT Alert 3D system, respectively (kappa coefficient, 0.884). The rate of drug resistance was 13.4% for at least one or more drugs, 6.0% for MDR-TB and 2.3% for XDR-TB. M-KIT plate and BacT 3D Alert 3D system were comparable in drug susceptibility testing for isoniazid (97.7%; kappa coefficient, 0.905) and rifampin (98.6%; kappa coefficient, 0.907). Antibiotic resistance was observed using M-KIT plates for 24 of the total 29 Mtb isolates (82.8%). Conclusion: The liquid culture system showed greater reduction in the culture period, as compared with LJ medium; however, drug susceptibility testing using M-KIT plates was advantageous for simultaneous testing against multiple drug targets.

• 한국산학기술학회논문지
• /
• 제12권11호
• /
• pp.5012-5018
• /
• 2011

1965년부터 1995까지 결핵의 전국적인 조사에 의하면, 한국에서 결핵의 발생과 약제 내성율은 감소하였지만 다재약제내성율의 유행은 여전히 심각한 문제이다. 본 연구의 목적은 2001 년부터 2008까지 대전에 약제 내성율과 성향을 조사하였다. 총 581건의 약제감수성 검사가 수행 되었으며 이중 하나이상의 항결핵제의 약물에 내성이 있는 경우가 104건(17.9%)이였고 적어도 INH에 내성이 있는 경우가 68건 (11.7 %), RFP내성이 41건(7.1 %)이였다. 단일 약제 내성율은 37건 (6.4 %) 분리되었으며 이들 중 INH에서 18건 (3.1 %) RFP에서 5건(0.9 %)이 분리 되었다. 적어도 isoniazid와 refampin에 내성인 다제내성결핵은 35건(6.0 %)에서 발견되었다. 그리고 MDR - TB와 관련된 나이요인에 40-60 세 포함되었다. 폐결핵의 약제내성율, 특히 MDR - TB의 약제내성율은 공공의료분야보다 민간 병원의 초치료 환자에서 높게 나왔다. 초기 약제내성은 일반적이며 약제감수성 검사는 이전 결핵 치료를 받지 않은 폐결핵 환자에 대해 유용성이 있다. 내성의 확산과 증가를 줄이기 위해 MDR - TB의 조기 진단과 함께 향상된 제어프로그램이 필요하다. 다제내성율은 여전히 한국에서 문제이다. 독립적으로 혹은 공공의료분야와 공동으로 다제내성율을 감소하기 위한 노력이 필요하다.