• Journal of Microbiology
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• 제42권2호
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• pp.80-86
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• 2004

Pulsed-field gel electrophoresis (PFGE) typing was applied to the epidemiological investigation of 21 Candida tropicalis isolates collected from urine specimens of 11 patients and one healthcare worker, in an intensive care unit (ICU) over a 4-month period. Seventeen epidemiologically unrelated strains from 14 patients were also tested to determine the discriminatory power of PFGE. PFGE typing consisted of electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG), using two restriction enzymes (BssHII and SfiI). The EK pattern was the same in all 38 isolates, while REAG using SfiI separated the isolates into nine types. However, 16 different PFGE types were iden-tified by REAG with BssHII, and the same results were obtained when the results of both REAG tests were combined. In serial urinary isolates from 10 patients, all strains from each patient had the same PFGE pattern. While the epidemiologically unrelated strains from 14 patients consisted of 13 different PFGE types, the 20 isolates from the 11 ICU patients fell into only two PFGE types (types Cl and C2), and these apparently originated from the two different outbreaks. All strains of type Cl (n = 12) were isolated from six patients, between November 1999 and January 2000, and all of the type C2 strains (n=8) were isolated from five patients, during January and February 2000. This study shows two con-secutive clusters of C. tropicalis candiduria in an ICU, defined by PFGE typing, and also demonstrates that a PFGE typing method using BssHII is perhaps the most useful method for investigating C. tropi-calis candiduria.

• Applied Biological Chemistry
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• 제37권1호
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• pp.56-63
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• 1994

오이모자이크 바이러스(CMV)의 외피단백질 유전자의 일정한 염기서열을 보유하는 부분과 CMV RNA에 대항한 헤머헤드(hammerhead) 구조의 ribozyme을 만드는 올리고뉴클 레오타이드(oligonucleotide, nt)를 DNA 합성기를 이용하여 제조하였다. 올리고뉴클레오타이드의 양쪽가닥을 서로 합친 후 제한효소 BamHl과 SacI으로 처리하여 플라스미드 pBS SK(+)에 삽입하였고 CMV 기질과 ribozyme 클론의 염기서열을 결정하여 확인하였다. 기질과 ribozyme 클론 $1\;{\mu}g$ BssHII이나 SspI으로 처리한후 T7 RNA 합성효소를 이용하여 튜브내에서 전사반응을 실시하였다. 제한효소 BssHII를 처리한 경우 만들어진 기질 RNA의 크기는 176 nt 였는데 50 nt의 CMV RNA 염기, 6 nt의 Xbal 제한효소 염기, 120 nt의 벡터에서 비롯된 염기를 포함한다. Ribozyme RNA의 크기는 164 nt인데 38 nt의 ribozyme 염기부분과 그외는 기질의 것과 같은 염기를 포함한다. CMV 기질 RNA는 ribozyme RNA에 의하여 특이적으로 절단되어 96 nt와 80 nt 두개의 조각을 만들었다. 이러한 특이적 절반은$37^{\circ}C$ 보다 $55^{\circ}C$에서 더 빠르게 일어났다. SspI으로 처리한 경우 만들어진 기질 RNA(2234 nt)도 역시 위치 ribozyme에 의해 두조각으로 절반되었으며 SspI 처리 후 만들어진 ribozyme RNA(2222 nt)에 의해서 특이적으로 절단되었다.

• 대한미생물학회지
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• 제35권2호
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• pp.191-201
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• 2000

The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with THP-1 and HSV-2 standard strain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of THP-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and THP-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, Band BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum form respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 47.6%) HSV was detected singly or mixed infection and 19 of those cases were HSV-2 and 1 case was THP-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, Band BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.

• Clinical and Experimental Pediatrics
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• 제53권3호
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• pp.432-436
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• 2010

일과성 신생아 당뇨병은 6번 염색체의 부친 이체성, 부친으로부터 유래한 염색체 6q24의 중복, ZAC 또는 HYMAI 유전자의 CpG 섬의 메틸화 결함에 의해 야기된다. 저자들은 고혈당, 거설증, 자궁내성장지연으로 발현되어 부친으로부터 유래된 HYMA1 유전자만을 보인 일과성 신생아 당뇨병 1례를 경험하였다. 생후 18일된 여아가 거설증과 반복되는 고혈당으로 입원하였다. 거설증과 함께 큰 대천문, 작은 턱, 두드러진 눈을 보였으며 혈중 포도당 농도는 200-300 mg/dL로 유지되다가 입원 2일 후부터 인슐린 투여 없이도 정상 범위로 유지되었다. 모체로부터 유래된 메틸화된 대립유전자 유무를 확인하기 위하여 말초 혈액으로부터 genomic DNA를 추출하여 bisulfite를 처리한 후, 메틸화 특이 중합 효소 연쇄 반응으로 HYMAI 유전자의 일부분을 증폭시키고, 제한 효소 BssHII를 이용한 제한 효소 절편 길이 다형성 (restriction fragment length polymorphism, RFLP) 분석을 이용하여 HYMAI 유전자의 메틸화 여부를 확인한 결과, HYMAI 유전자의 메틸화 결함을 보여 부친에서 유래된 HYMAI 유전자만을 갖고 있음을 확인하였다. HYMAI 유전자의 메틸화 검사를 통해 6번 염색체의 각인된 유전자가 증명되었으며 메틸화 결함으로 인해 일과성 신생아 당뇨병이 발생하였다.

• Journal of Microbiology
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• 제43권6호
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• pp.546-554
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• 2005

We assessed the genetic relations and epidemiological links among bloodstream isolates of Candida albicans, which were obtained from a university hospital over a period of five years. The 54 bloodstream isolates from the 38 patients yielded 14 different karyotypes, 29 different patterns after digestion with SfiI (REAG-S), and 31 different patterns after digestion with BssHII (REAG-B) when analyzed using three different pulsed-field gel electrophoresis (PFGE) typing methods. In 11 patients with serial blood stream isolates, all strains from each patient had the same PFGE pattern. The dendrograms for all of the strains revealed that the distribution of similarity values ranged from 0.70 to 1.0 in the REAG-S patterns, and from 0.35 to 1.0 in the REAG-B patterns. Overall, the combination of the three different PFGE methods identified 31 distinct types, reflecting the results obtained using the REAG-B alone different. different Five PFGE types were shared among 22 isolates from 12 patients. These types of strains were more frequently associated with central venous catheter-related fungemia than the other 26 type strains $(92\%\;versus\;31\%;\;P<0.005)$. Of five PFGE types, four isolates were determined to be epidemiologically related: each of these types was primarily from two or three patients who had been hospitalized concurrently within the same intensive care unit. Our results suggest that the REAG-B constitutes perhaps the most useful PFGE method for investigating C. albicans candidemia and also shows that a relatively high proportion of C. albicans candidemia may be associated with exogenous acquisition of clonal strains.