• Korean Journal of Veterinary Research
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• v.52 no.1
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• pp.39-43
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• 2012

A multiplex PCR was developed for the simultaneous detection and differentiation of Mycoplasma (M.) hyopneumoniae and M. hyorhinis in clinical samples. Improved sensitivity is advantage of this technique over the previously reported multiplex assay. It was capable of detecting as little as 125 fg genomic DNA from M. hyopneumoniae and 62.5 fg genomic DNA from M. hyorhinis. Application of this multiplex PCR method to field isolates showed that M. hyopneumoniae and M. hyorhinis were present in 29% (107 of 370) of lung specimens and no mycoplasmas were detected in 56% (208 of 370) of the slaughtered pigs' lungs. At the farm level, M. hyopneumoniae and M. hyorhinis were detected in 34 of 36 (94.4%) randomly selected farms. We conclude that this assay would prove itself a value tool for monitoring these mycoplasmal infections and both M. hyopneumoniae and M. hyorhinis have been widely spread in swine herds of Korea.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.265-270
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• 2009

An oligonucleotide array was developed to detect and genotype mollicutes based on the internal transcribed spacer (ITS) sequence. The results of the assay were compared with those of a PCR-RFLP assay. The proposed oligonucleotide array containing 5 genus- and 23 species-specific probes was able to detect Mycoplasma species, including M. penetrans and M. spermatophilum, that were not detected by the PCR-RFLP assay. Therefore, the results demonstrated that the proposed oligonucleotide array was effective for the detection and discrimination of 23 species, including an acholeplasma, 21 mycoplasmas, and a ureaplasma, and showed promise as a countermeasure to ensure that biological products are safe and of good quality.

• Korean Journal of Veterinary Service
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• v.46 no.1
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• pp.15-27
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• 2023

Bordetella (B.) bronchiseptica, Mycoplasma (M.) cynos, and M. canis are the major bacterial pathogens that cause canine infectious respiratory disease complex (CIRDC). In this study, we developed a triplex real-time polymerase chain reaction (tqPCR) assay for the differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with a detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra- and inter-assay variations of less than 1%. The diagnostic results of the assay using 94 clinical samples from household dogs with CIRDC clinical signs, the prevalence of B. bronchiseptica, M. cynos, and M. canis was 22.3%, 18.1%, and 20.2%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported qPCR assays. The dual infection rate of B. bronchiseptica and M. cynos, B. bronchiseptica and M. canis, and M. cynos and M. canis was 5.3%, 7.4%, and 3.1%, respectively. Moreover, the triple infection rate of B. bronchiseptica, M. cynos, and M. canis was 2.1%. These results indicate that coinfections with B. bronchiseptica, M. cynos, and M. canis have frequently occurred in the Korean dog population. The newly developed tqPCR assay in the present study will be a useful tool for etiological and epidemiological studies on these three CIRDC-associated bacterial pathogens. The prevalence and coinfection data revealed through this study will contribute to expanding knowledge on the epidemiology of CIRDC in the recent Korean dog population.

• Korean Journal of Veterinary Research
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• v.42 no.2
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• pp.225-229
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• 2002

We report that mycoplasma organisms from lung tissues of slaughter pigs were identified to genes fragments with references use of nested-PCR technique(nPCR). Seven strains of mycoplasma species were isolated from 70 lung tissues. The organisms were detected by in vitro amplification of 16S rRNA and 23S rRNA genes. Nucleotide sequences of the spacer between 16S and 23S in the ribosomal RNA operons of mycoplasma were identified by the analysis of products from the nested PCR. Four common PCR primers, MhF1, MhF2 MhR1 and MhR2, were designed by analysis between these sequences by first amplified with F1, R1 and second with F2, R2, respectively. Specific amplification of the spacer region for reference strains of M. hyopneumoniae, M. hyorhinis, M. flocculare were confirmed by first round of PCR in which the traduced fragments of 690bp, 460bp, 630bp. But amplications of second round was changed to 240bp, 210bp, 230bp, respectively. Three different strains (M. hyopneumoniae:4, M. hyorhinis:2, M. flocculare:1) were detected by the nested-PCR technique. The results suggest that the detection of swine mycoplasma by n-PCR can be analyzed the nucleotide sequences between rRNA operons and homology study.

• Korean Journal of Veterinary Service
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• v.39 no.2
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• pp.101-105
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• 2016

The present study investigated serological and molecular prevalence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infection in unvaccinated broiler breeder farms in Jeonbuk providence. An enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) had been used to determine antibody titers against MG and MS, and genome of these pathogens, respectively. Seventy five percent of farms were seropositive for MG and 94% of farms were seropositive for MS. In addition, the rate of antibody positive flocks against MG were 65.3% (32/49), while the rate of positive flocks against MS were 84.2% (80/95). The geometric mean antibody titers were $802.2{\pm}626$ and $27,726.7{\pm}2426$ against MG and MS, respectively. Interestingly, none of samples was positive for MG genome by PCR, while 94% (farms), 82% (flocks) and 62.6% (broiler breeder) were positive for MS genome by PCR. These findings suggest that the prevalence of MG or MS infection could be higher than expected. Thus, strict prevention program including vaccination and environmental sanitation should be implemented to avoid disease transmission from breeder to broilers as well as transmission among broilers.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2003.10a
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• pp.29-29
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• 2003

Mycoplasma hyopneumoniae is among the most prevalent and important infectious agents associated with porcine respiratory disease complex. The airway dagame caused by M. hyopneumoniae adversely affect the pulmonary host defense mechanisms and may lead to secondary bacterial infections. Culture is considered to be the "gold standard" for diagnosis but this is a very slow and labor-intensive procedure. Isolation of M. hyopneumoniae is complicated by its fastidious nature and extremely slow growth. Thirty days of incubation may be necessary to detect the organism in primary broth cultures. The purposes of the study were (ⅰ) to develop nested PCR for the detection of M. hyopneumoniae for the detection of M. hyopneumoniae DNA in the formalin-fixed, paraffin-embedded lung tissues from experimentally and naturally infected pigs and (ⅱ) to compare the utility of nested PCR with in situ hybridization. (omitted)

• Clinical and Experimental Pediatrics
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• v.52 no.3
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• pp.315-323
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• 2009

Purpose : This study aimed to perform a systematic review of the reports on Mycoplasma pneumoniae pneumonia in the last 30 years (1980-2006) to investigate the intervals between outbreaks, change in the peak incidence age, and diagnostic methods. We also aimed to validate the proper diagnostic criteria for M. pneumoniae pneumonia. Methods : We reviewed 62 original articles on M. pneumoniae pneumonia in Korean children. We analyzed the annual or seasonal variation, study areas, patient age, journal names, and the date of each report. Further, we checked the methods and criteria used for the diagnosis of M. pneumoniae pneumonia. We also confirmed the proper mycoplasma antibody cutoff using the mycoplasma IgM titer as the gold standard. Results : In the last 30 years, epidemic outbreaks of M. pneumoniae pneumonia occurred every 3 years, except in 1993-1994 and 1996-1997. Seasonal variations were also present and were most prevalent in October and November. The number of preschool children, especially those aged 3 years or younger, with M. pneumoniae pneumonia has increased (P<0.05). The mycoplasma antibody titer of 1:640 or greater was appropriate for diagnosing M. pneumoniae pneumonia, with an acceptable sensitivity and specificity of detection. Conclusion : We analyzed the results of studies on M. pneumoniae pneumonia in Korean children during the last 30 years. Infection in younger children is increasing, and further research is required to reveal the major cause of the changing epidemics.

• Journal of Korean Society of Forest Science
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• v.67 no.1
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• pp.28-30
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• 1984

Distribution of mycoplasma-like organisms (MLO) in the phloem tissue of witches'-broom infected jujube trees was investigated by fluorescence microscopy applying new fluorochrome DAPI (4'-6-diamidino-2-phenylindole, 2HCl). MLO were detected from the phloem of leaf and stem sections of diseased branches exhibiting typical witches'-broom symptom but not detected from those of symptomless branches of an infected tree. MLO were also present in the healthy looking leaves frequently found in the diseased shoots. Fluorescence microscopy revealed the presence of MLO in the phloem of root sections of infected trees.

• Korean Journal of Veterinary Service
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• v.45 no.4
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• pp.305-316
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• 2022

Bordetella (B.) bronchiseptica, Mycoplasma (M.) felis, and Chlamydia (C.) felis are considered as main bacterial pathogens of feline upper respiratory tract disease (URTD). In this study, a new triplex quantitative real-time polymerase chain reaction (tqPCR) assay was developed for the rapid and differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with the detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 1%. Based on the diagnostic results of the assay using 94 clinical samples obtained from cats with URTD signs, prevalence of B. bronchiseptica, M. felis, or C. felis was 10.6%, 36.2%, or 6.4%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported monoplex qPCR assays. The dual infection rates for B. bronchiseptica and M. felis or M. felis and C. felis was 2.1% or 3.2%, respectively. These results indicated that M. felis has been widely spread, and its co-infection with B. bronchiseptica or M. felis has been frequently occurred in Korean cat population. The developed tqPCR assay will serve as a promising tool for etiological and epidemiological studies of these three bacterial pathogens and the prevalence data obtained in this study will contribute to expanding knowledge about the epidemiology of feline URTD in Korea.

• Korean Journal of Veterinary Service
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• v.31 no.1
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• pp.71-77
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• 2008

Today swine respiratory disease is one of the most important diseases because of its economic losses and severe infection nationwide, and swine society as well as veterinary service are trying to prevent the diseases in Korea. This study would like to obtain some information useful for the control of the diseases. A total of 174 lung specimens with lesion consisted of 3 sorts; 60 were collected from nursey pigs requested for diagnostic service from March of 2006 to October of 2007, 58 finishing pigs and 56 sows were selected from slaughterhouse from September to November 2007. In the detection test of pathogens by PCR, porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae were positive in 95.4%, 31.6%, and 20.1%, respectively. Double infection rate with PCV2 and PRRS was 30.4%, PCV2 and M hyopneumoniae was 19.5%, triple infection with PCV2, PRRS and M hyopneumoniae was 5.7%, respectively.