• Pediatric Infection and Vaccine
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• 제26권2호
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• pp.81-88
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• 2019

목적: Mycoplasma pneumoniae 폐렴은 학동기 소아와 청소년의 지역사회 획득 폐렴 중 가장 흔한 원인으로, 조기에 원인 진단이 가능하다면 적절한 항균 요법을 결정하는데 도움이 된다. 본 연구는 하기도 감염 소아의 호흡기 검체에서 M. pneumoniae를 검출하기 위한 신속 항원 검사 방법의 진단적 가치를 평가하고자 하였다. 방법: 2010년 8월부터 2018년 8월까지 하기도 감염으로 서울대학교 어린이병원에서 응급실 또는 입원 치료를 받은 소아로부터 채취한 비인두 흡인물 중 M. pneumoniae 배양 검사를 시행한 후 $-70^{\circ}C$ 초저온냉동고에 보관되어 있는 검체 215개를 선정하였다. 비인두 흡인물 검체를 실온에서 해동하고 면역크로마토그래피를 이용한 Ribotest $Mycoplasma^{(R)}$를 시행한 후 두 명의 검사자가 결과를 판독하였다. 검사를 시행하는 자와 판독하는 자는 배양 검사 결과를 모르는 상태에서 검사를 진행하였다. 결과: 총 215개의 비인두 흡인물 검체 중 M. pneumoniae가 배양 양성인 검체는 119개, 배양 음성인 검체는 96개였다. M. pneumoniae가 배양 양성인 119개 중 74개(62.2%)가 Ribotest $Mycoplasma^{(R)}$ 검사 결과 양성이었고, 배양 음성인 96개 중 92개(95.8%)가 Ribotest $Mycoplasma^{(R)}$ 검사 결과 음성이었다. 배양 검사 결과를 기준으로 평가한 Ribotest $Mycoplasma^{(R)}$의 민감도는 62.2%(74/119, 95% 신뢰구간, 53.5-70.9%)이었으며, 특이도는 95.8% (92/96, 95% 신뢰구간, 91.8-99.8%)이었다. 또, 양성 예측도는 94.9% (74/78, 95% 신뢰구간, 90.0-99.8%)이었으며, 음성 예측도는 67.2% (92/137, 95% 신뢰구간, 59.3-75.0%), 그리고 일치도 77.21% (166/215, 95% 신뢰구간, 71.6-82.8%)를 보였다. 결론: 본 연구 결과, 신속 항원 검출법인 Ribotest $Mycoplasma^{(R)}$ 검사결과가 양성인 경우는 M. pneumoniae 배양 양성과의 일치도가 매우 높아서 M. pneumoniae 감염의 진단에 유용하였다. 그러나, Ribotest $Mycoplasma^{(R)}$ 검사결과가 음성인 경우의 약 1/3은 M. pneumoniae 배양 양성인 검체이었으므로, 음성 검사 결과에 대한 해석은 주의하여야 한다.

• 한국동물위생학회지
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• 제22권1호
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• pp.9-13
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• 1999

Mycoplasmal pneumonia of swine(MPS) cause by Mycoplasma hyopneumoniae has been recognized as a serious impediment to swine production due to chronic respiratory disorder which result in the weight loss and decreased feed conversion. The disease causes a great economic losses in pig industry by characterizing with high morbidity, low mortality, growth retardation and low feed efficiency. The present study was conducted to investigate the titers of antibody against M hyopneumoniae from the regional and seasonal groups of the slaughtered pigs by enzyme-linked immunosorbent assay(ELISA). The result have shown that the average seropositive rate of M hyopneumoniae infection was 84.6% . The regional seropositive rate in Korea showed 87.4% in Kyonggj, 83.4n in Kangwon, 89.2% in Chungnam and 77.6% in Chungbuk area, respectively. Also the seasonal seropositive rate was appeared as 78.6% in spring,90.1% in summer, 76.9% in autumn and 83.8% in winter, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.84-91
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• 2000

Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

• 한국가금학회지
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• 제46권4호
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• pp.249-253
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• 2019

닭 마이코플라즈마병은 전세계적으로 양계산업에 문제시 되고 있는 난계대 질병으로 병아리 품질 및 사육성적에 영향을 미친다고 알려져 있다. 본 연구는 닭 마이코플라즈마병에 대한 백신 접종을 실시하지 않은 계열화 회사의 육용 종계군을 대상으로 정기 채혈을 통해 혈청검사를 실시한 후 감염율을 확인하였고, 조사계군에서 생산된 육계 병아리에 대한 사육성적을 확인하고자 하였다. 육용종계 닭 마이코플라즈마 감염율과 그에 따른 후대병아리의 사육성적을 연도별로 확인한 결과, 종계군의 감염율이 낮아짐에 육계의 사육성적이 높아진다는 상관관계를 확인하였다. 이러한 결과를 토대로 비추어 보았을 때, 닭 마이코플라즈마병의 감염 유무는 생산된 초생추의 품질과 사육농장 성적 영향에 미치는 여러 요소들 중 하나라고 판단할 수 있다.

• 한국산림과학회지
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• 제80권2호
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• pp.232-236
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• 1991

형광색소(螢光色素) berberine sulphate의 DNA 및 RNA-결합성질(結合性質)을 이용(利用)한 형광현미경기법(螢光顯微鏡技法)의 마이코플라스마(mycoplasma-like organism ; MLO) 감염진단(感染診斷)에 대(對)한 효용가치(效用價値)를 조사(調査)하였다. 이병(罹病) 대추나무, 오동나무, 뽕나무 및 일일초(日日草)의 조직절편(組織切片)을 berberine sulphate로 염색(染色)하여 형광현미경(螢光顯微鏡)으로 관찰(觀察)하였던 바, 사부조직(篩部組織)에서 MLO-특이형광반응(特異螢光反應)이 나타났으나, 건전조직(健全組織)의 사부(篩部)에서는 특이형광(特異螢光)이 관찰(觀察)되지 않았다. 이와 같은 berberine sulphate의 조직염색법(組織染色法)은 목본(木本) 및 초본식물(草本植物)의 MLO감염(感染)을 정확(正確)하게 진단(診斷)하는데 매우 유용(有用)한 방법(方法)임을 보여 주었으며, 사용(使用)이 간단(簡單)하고 비용(費用)이 저렴(低廉)하므로 대량(大量)의 시료(試料)에 대한 신속(迅速)한 검정(檢定)에 이용(利用)될 가능성(可能性)을 보여 주었다.

• 한국동물위생학회지
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• 제38권2호
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• pp.101-106
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• 2015

Murine mycoplasmosis, caused by Mycoplasma (M.) pulmonis, is a prominent disease in rodent animals. The aim of this study was to develop a sensitive and specific PCR assay to detect M. pulmonis in animals and to assess the suitability of this assay for the detection of mycoplasmal infection in rats experimentally infected with M. pulmonis. A new PCR assay using the M. pulmonis-specific primer pairs MPul-F and MPul-R was developed. The primers and probe for the assay were designed from regions in the 16S rRNA gene that are unique to M. pulmonis. The novel PCR assay was very specific and sensitive for M. pulmonis, detecting the equivalent of 5 pg of target template DNA. It detected only M. pulmonis and no other Mycoplasma species or other bacterial species. The newly developed PCR assay also effectively detected M. pulmonis infection in rats. These results suggest that this PCR assay using M. pulmonis-specific primer pairs of MPul-F and MPul-R will be useful and effective for monitoring M. pulmonis infection in animals.

• Journal of Microbiology
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• 제44권1호
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• pp.42-49
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• 2006

In this study, we describe our newly-developed sensitive two-stage PCR procedure for the detection of 13 common mycoplasmal contaminants (M. arthritidis, M. bovis, M. fermentans, M. genitalium, M. hominis, M. hyorhinis, M. neurolyticum, M. orale, M. pirum, M. pneumoniae, M. pulmonis, M. salivarium, U. urealyticum). For primary amplification, the DNA regions encompassing the 16S and 23S rRNA genes of 13 species were targeted using general mycoplasma primers. The primary PCR products were then subjected to secondary nested PCR, using two different primer pair sets, designed via the multiple alignment of nucleotide sequences obtained from the 13 mycoplasmal species. The nested PCR, which generated DNA fragments of 165-353 bp, was found to be able to detect 1-2 copies of the target DNA, and evidenced no cross-reactivity with the generated DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. The identification of contaminated species was' achieved via the performance of restriction fragment length polymorphism (RFLP) coupled with Sau3AI digestion. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the disagnosis of mycoplasmal contamination in cell culture systems.

• Bulletin of the Korean Chemical Society
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• 제29권1호
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• pp.153-158
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• 2008

A microchip-based capillary gel electrophoresis (MCGE) technique was developed for the ultra-fast detection and differentiation of Candidatus Mycoplasma haemominutum (Candidatus M. haemominutum, California strain) and Mycoplasma haemofelis (M. haemofelis, Ohio strain) in Korean feral cats through the application of programmed field strength gradients (PFSG) in a conventional glass double-T microchip. The effects of the poly (ethyleneoxide) (PEO) concentration and electric field strength on the separation of DNA fragments were investigated. The PCR-amplified products of Candidatus M. haemominutum (202-bp) and M. haemofelis (273-bp) were analyzed by MCGE within 75 s under a constant applied electric field of 117.6 V/cm and a sieving matrix of 0.3% PEO (Mr 8 000 000). When the PFSG was applied, MCGE analysis generated results 6.8-times faster without any loss of resolution or reproducibility. The MCGE-PFSG technique was also applied to eleven samples selected randomly from 33 positive samples. The samples were detected and differentiated within 11 s. The analysis time of the MCGE-PFSG technique was approximately 980-times faster than that using conventional slab gel electrophoresis.

• 대한수의학회지
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• 제60권3호
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• pp.109-116
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• 2020

This study aimed to develop a semi-nested polymerase chain reaction assay for the direct detection of Mycoplasma synoviae (M. synoviae) from clinical samples using three newly designed oligonucleotide primers specific to the variable lipoprotein haemagglutinin (vlhA) gene and differentiate M. synoviae field strains based on a nucleotide deletion or the insertion of the proline-rich repeat (PRR) region of the vlhA gene. The developed semi-nested polymerase chain reaction (PCR) assay revealed positive results in 12 out of 100 clinical samples collected from chickens showing lameness and joint swelling. Six positive samples were selected randomly for sequencing, and sequence analysis revealed 96.3-100% nucleotide identities compared to the reference sequences. Phylogenetic analysis showed that sequences of the strains in this study were closely related to WVU1853 (Spain), CK.MS.UDL.PK.2014.2 (Pakistan), and F10-2AS (USA) strains, but they were distinct from the M. synoviae-H vaccine strain sequence. M. synoviae obtained from these samples were identified as types A and C with a length of 38 and 32 amino acids, respectively. These results indicated that the specific and sensitive semi-nested PCR could be a useful diagnostic tool for the direct identification of clinical samples, and the sequence analysis of the partial vlhA gene can be useful for typing M. Synoviae.

• 한국식물병리학회지
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• 제1권1호
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• pp.12-16
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• 1985

마이코플라스마에 감염된 대추나무, 뽕나무 및 일일초의 조직절편을 형광염색소인 DAPI(4'-6-diamidino-2-phenylindole$\cdot$2HCl), aniline blue 그리고 quinacrine(quinacrine mustard dihydrochloride)으로 염색하여 형광현미경하에서 관찰함으로서 이들 형광염색소의 마이코프라스마 감염진단에의 효용가치를 비교 조사하였다. 이병식물의 줄기절편의 절부에서 특이형광반응이 나타났으나 건전식물의 조직절편에서는 특이형광반응이 관찰되지 않음으로써, DAPI, anilinc blue 및 quinacrine 등의 형광염색소에 의한 조직염색법은 대추나무, 뽕나무 및 일일초의 마이코플라스마 감염을 식속 정확하게 진단하는 데 매우 유용한 방법임을 보여주었다. 한펀, 이들 형광염색소 중에서는 DAPI가 가장 효과적이었다.