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

• 대한수의학회지
• /
• 제26권2호
• /
• pp.283-292
• /
• 1986

Isolation and identification of Mycoplasma were performed to clarify Mycoplasma infection of mice fed by conventional feeding at two ($K_1$, $K_2$) institutes in Korea. The twenty mice to be tested were randomly sampled from each of 10 breeding colonies in respective institute. Identification of the Mycoplasma strains isolated from the nasal cavity, lung and synovia of mice was made according to the morphology of colonies, biological and biochemical properties with special reference to M. pulmonis, M. arthrotodis and M. neurolyticum. In addition, growth inhibition test was performed using hyperimmune rabbit antisera to the strain PG-22 of M. pulmonis, the strain PG-6 of M, arthritidis and the strain PG-28 of M. neurolyticum and also differentiation of isolates from L-form bacteria was dont by Dieses staining and culture method with passage of the isolates on liquid media eliminated antibacterial drug. On the other hand, a total of 13 strains out of the 44 isolated M. pulmonis from mice was investigated for their susceptibility against 16 antibiotics in vitro. The antibiotic sensitivity test was made using $3{\times}10^4$ organisms/0.3ml on each plate(90mm diameter) with antibiotic mono-or tri-disk. The results obtained are summarized as follows: 1. Out of 20 mice from 10 breeding colonies in Kl institute, mycoplasma-like strains from the nasal cavity of 16 mice(80%) and from the lung of 8 mice(40%) were isolated, while out of 20 mice in K2 institute, M-like strains were isolated from the nasal cavity of 14 mice(70%) and from the lung of 6 mice(30%). However, no mycoplasma-like organisms were isolated from the synovia of the 40 mice examined. All the 44 strains isolated were identified as the organisms of M. pulmonis. 2. Out of the 16 antibiotics tested, penicillin, oleandomycin and bacitracin showed no activity against all the 13 M. pulmonis strains. On the contrary, lincomycin, clindamycin, chloramphenicol, tetracycline, minocycline, kanamycin, gentamycin and tobramycin showed high activity with three different antibiotic concentration of tridisk, but amikasin and spiramycin showed intermediate activity. Other antibiotics such as polymyxin B and colistin showed low activity, while erythromycin showed lower activity than others.

• 대한수의학회지
• /
• 제29권4호
• /
• pp.517-523
• /
• 1989

This study was undertaken to establish reliable diagnostic-procedures for the microbiological monitoring of laboratory animals. Murine(mice and rats) antibodies against hemagglutinating virus of Japan(HVJ), mouse hepatitis virus(MHV) and Mycoplasma pulmonis(Mp) were detected sensitively and specifically in experimentally and naturally infected animals' sera by an indirect enzyme-linked immunosorbent assay(ELISA), using urease conjugated antimurine immunoglobulin. The sensitivity and specificity of the complement fixation test which has been apllied widely for serodiagnosis of HVJ, MHV and Mp infections were apparently lower than those of ELISA. From these results, the ELISA was found to be available for the serodiagnosis of HVJ, MHV and Mp infections in mice and rats.

• 환경위생공학
• /
• 제1권1호
• /
• pp.69-79
• /
• 1986

랫트와 마우스에 M. Pulmonis의 감염률을 조사하기 위해 마우스 310수 랫트 330수를 사용하여 이 균에 의한 여러가지 친화성조직중 가장 높게 감염을 일으키는 부위와 각 채취부위별로 검체의 배양검사 및 보체결합 반응에 따른 특이항체 양성률등의 실험을 수행하여 다음 결론을 얻었다. 1. 마우스와 렛트에서 M. Pulmonis의 양성률이 높은 부위는 마우스에서 구강, 안, 비강순이고 랫트에서 비강, 기관지, 구강 순으로 나타냈다. 2. M. Pulmonis 양성률은 A사육장의 마우스 50수중 6수$(12\%)$이며, B사육장에서는 8수$(16\%)$이었다. 한편 랫트에서 60수중 33수$(55\%)$와 42수$(70\%)$로 각각 나타냈다. 3. 랫트와 마우스에서 채취한 혈청으로 보체결합 반응을 통하여 얻은 특이적인 항체가가 1:5(< 1:5)보다 낮은 경우는 마우스 100수중 73수$(73\%)$이고 1:5 보다 높은 항체가를 보인 경우에는 24수$(24\%)$ 이었으며 나머지 3수$(3\%)$는 항보체기능으로 측정하지 못했다. 한편 랫트에서 1: 5보다 낮은 항체가를 보인 경우에는 총 120수중 42수$(35\%)$ 이고 1: 5보다 높은 경우 73수$(61\%)$의 양성율을 보였으며 나머지 $6\%$는 항보체 기능으로 측정이 안되었다.

• 대한수의학회지
• /
• 제41권4호
• /
• pp.579-582
• /
• 2001

SHR 랫드 4-21 개월령 colony에서 혈청 및 조직학적으로 정기적인 health monitoring을 실시하던 중 ELISA test에서 21 개체 중 12 개체에서 Mycoplasma pulmonis에 대한 항체가 검출되었다. Mycoplasma 양성 개체군의 부검 소견에서 기관지, 세기관지 주변 림프절의 증식과 전형적인 폐 기관지의 병변 소견을 보였다. 혈청학적 검사와 조직학적 소견에서 SHR 랫드 군의 자연적인 Mycoplasm 감염 예로 진단되어 보고하는 바이다.

• 대한수의학회지
• /
• 제40권3호
• /
• pp.611-625
• /
• 2000

For the improvement of quality control of laboratory mouse, we investigated the environmental condition, histopathological findings and serological test using ELISA to mouse hepatitis virus(MHV), Mycoplasma pulmonis(MP), Clostridium piliforme(TZ) and Sendai virus (HVJ) of ICR, C57BL/6, CBA and C3H/He mice that were supplied from conventional laboratory animal facility. 1. The ammonia concentration of facility was below the recommended concentration, 15ppm, by the KNIH, and the room temperature($21{\sim}23^{\circ}C$) and relative humidity(40~60%) was optimum range recommend by the Ministry of Health and Welfare, respectively. 2. The incidence rate of inapparent disease was 86.6% and the major findings in the liver were vacuolar degeneration with nucleic pleomorphism. The lung was shown the thickening of alveolar wall and interstitial pneumonia with congestion. The kidney and spleen were observed the mild congestion and extramedullary hematopoiesis, respectively. 3. The positive reaction rates against MHV and MP in serological test was 97.9% and 37.5%, respectively but HVJ and TZ were negative. These results suggest that laboratory mice could be infected with MHV and MP under conventional environments. Therefore we recommend to select thoroughly inapparent infected mice and to convert conventional system into SPF facility as soon as possible.

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

• 생명과학회지
• /
• 제17권2호통권82호
• /
• pp.279-285
• /
• 2007

위암 환자 30명과 결장암 환자 30명의 암 조직과 그 주위의 정상 조직을 구분하고, 암 조직과 정상조직의 쌍을 찾지 못한 경우의 위암 조직 8개와 결장암 조직 10개의 각 조직에서 Mycoplasma DNA의 존재 유무를 PCR법으로 확인하고 PCR산물에서 DNA의 염기서열을 분석하여 Mycoplasma DNA 서열과 비교함으로써 Mycoplasmas를 검색한 결과 다음과 같은 결론을 얻었다. 위암 조직과 그 주위 조직 30개 중에서 Mycoplasma DNA가 검출된 것은 각각 13개(43.3%)와 18개(60%)이었으며, 결장암 조직과 그 주위 조직 30개 중에서 Mycoplasma DNA가 검출된 것은 각각 12개(40%)와 15개(50%)이었다. Mycoplasma DNA가 검출된 위암 조직에서 mycoplasma 균종의 분리빈도는 M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. conjunctivae의 순이었다. Mycoplasma DNA가 검출된 위암 조직 주변의 정상조직에서 mycoplasma 균종은 M. facium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. bovigenitalium와 M. pulmonis의 순이었다. Mycoplasma DNA가 검출된 결장암 조직에서 mycoplasma 균종의 분리 빈도는 M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae), M. synoviae M. bovigenitalium, M. gallinarum, M. moatsii의 순이었다. Mycoplasma DNA가 검출된 결장암 조직 주변의 정상조직에서 mycoplasma 균종은 M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. bovis, M. opalescens, M. bovigenitalium, M. gallinarum와 M. moatsii의 순이었다. 이상의 결과에서 위암 및 결장암 조직 보다 암 주위의 정상 조직에서 Mycoplasma DNA의 검출율이 높았으며, 검출된 Mycoplasma 균종도 암 조직과 정상 조직에서 차이가 없었으므로 이들 암과 Mycoplasma와는 상관관계가 없는 것으로 생각된다.