• 제목/요약/키워드: PCR diagnosis

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Efficacy of Induced Sputum for the Diagnosis of Pulmonary Tuberculosis in Adults Unable to Expectorate Sputum

  • Park, Jae Seuk
    • Tuberculosis and Respiratory Diseases
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    • 제78권3호
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    • pp.203-209
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    • 2015
  • Background: Induced sputum (IS) has been used to collect airway secretions in subjects who have inadequate sputum production. The aim of this study was to investigate the efficacy of IS for the diagnosis of pulmonary tuberculosis (PTB) in adults unable to expectorate sputum. Methods: Medical records of 39 PTB patients who underwent IS due to absence of spontaneous sputum production between January 2011 and March 2014 at a tertiary hospital in South Korea were reviewed. Results of acid fast bacilli smear, Mycobacterium tuberculosis culture and polymerase chain reaction assay for M. tuberculosis (TB-PCR) of IS specimens from these patients were analyzed. Clinical and high-resolution computed tomography (HRCT) characteristics were also analyzed to find characteristics associated with IS culture positivity. Results: Of the 39 IS specimens from PTB patients, 7 (17.9%) were smear positive and 31 (79.5%) were culture positive. Twenty-four IS specimens were tested for TB-PCR and 13 (54.2%) were positive on TB-PCR. Multivariate analysis showed that younger age (p=0.04) and presence of tree-in-bud appearance on HRCT (p=0.03) were independent predictors of IS culture positivity. Conclusion: IS is useful for the diagnosis of PTB in adults unable to expectorate sputum. Younger age and tree-in-bud appearance on HRCT were associated with IS culture positivity in these patients.

중합효소연쇄반응을 이용한 닭 종양성 질병의 감별진단에 관한 연구 (Differential diagnosis among Marek's disease, reticuloendotheliosis and avian leukosis by polymeras chain reaction)

  • 성환우;김선중
    • 대한수의학회지
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    • 제38권1호
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    • pp.101-106
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    • 1998
  • The present study attempted to apply polymerase chain reaction (PCR) to develop a rapid differential diagnosis among Marek's disease, reticuloendotheliosis and avian leukosis. The primers chosen to detect Marek's disease virus (MDV) flank the 132bp tandem direct repeat of the MDV genome. The primers selected for reticuloendotheliosis virus (REV) and avian leukosis virus (ALV) are based on proviral long terminal repeats of spleen necrosis virus and Rous-associated virus-2 genomes, respectively. The specific PCR products of MDV, REV and ALV were observed with each primer and the reaction was not cross-reacted among the viruses. MDV-specific DNA was also amplified from the MDV-induced lymphoma (MDCC-MSB1) but not from the REV-induced tumor and ALV-induced lymphoma (LSCC-1104B1). In addition, proviral DNA of REV from REV-induced tumor and proviral DNA of ALV from ALV-induced lymphoma were also amplified by REV-specific and ALV-specific PCRs, respectively. Therefore these three PCR methods may be used to rapidly differentiate among MDV, REV and ALV-associated tumors in diagnosis.

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Mycoplasma hyopneumoniae와 Mycoplasma hyorhinis 동시 감별진단을 위한 다중진단 중합효소반응 (Simultaneous diagnosis and differentiation of Mycoplasma hyopneumoniae and Mycoplasma hyorhinis infections by multiplex PCR)

  • 홍선화;이현아;김동우;김태완;김옥진
    • 한국동물위생학회지
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    • 제37권4호
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    • pp.247-252
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    • 2014
  • The economic impact of swine mycoplasma infection is high. An accurate diagnosis is often difficult and time consuming. We report the development and validation of an effective multiplex polymerase chain reaction (PCR) assay that detects Mycoplasma (M.) hyopneumoniae and M. hyorhinis. The multi detection of M. hyopneumoniae and M. hyorhinis primer set were employed to detect mycoplasma species and typing of the species was performed on the basis of sequence analysis of the PCR product. The target nucleic acid fragments were specifically amplified by M. hyopneumoniae and M. hyorhinis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were used to evaluate the specificity of the multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. hyopneumoniae and M. hyorhinis.

근이양증 가계에서의 PEP-PCR을 이용한 착상전 유전자진단 (Preimplantation Genetic Diagnosis Using Primer Extension Preamplification in Duchenne/Becker Muscular Dystrophy(DMD/BMD) Families)

  • 최수경;이은호;이호준;전진현;강인수;백은찬;류현미;전종영
    • Clinical and Experimental Reproductive Medicine
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    • 제23권1호
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    • pp.109-114
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    • 1996
  • General PCR technique alone has a limitation for preimplantation genetic diagnosis(PGD) using single blastomere. Recntly developed primer extension preamplification(PEP) technology amplifies the whole genome and thus, simultaneous multiple locus analysis became possible. In this study, we report the efficacy of PEP-PCR for PGD in three muscular dystrophy carriers undergoing IVF-ET. A total of 37 blastomeres were obtained from 40 embryos at six to eight cell stage in three IVF cycles in two DMD and one BMD carriers. Whole genome from single blastomeres were amplified using I5-base oligonucleotide random primers. PCR amplified products of exon 45 in the dystrophin gene and alphoid X/Y loci for gender determination were analysed by 2% metaphor gel electrophoresis. A total of 37 PEP-PCR replicates from 37 single blastomeres from 40 embryos and 37 blanks were performed. We obtained the reliable results for exon 45 and alphoid X/Y. Transfer of female embryos and unaffected male embryo was attempted in three couples. Unfortunately, pregnancy was not achieved in these cases. PEP-PCR is a reliable and efficient PGD method in multiple locus analysis using single blastomere.

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파라핀 블록 PCR을 이용한 소 네오스포라 감염증의 진단법 확립 (Establishment of diagnositc method for bovine neosporosis by PCR using paraffin block)

  • 이중근;김재훈;김진현;이병천;황우석;윤희정;남호우;진영화;김대용
    • 대한수의학회지
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    • 제41권3호
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    • pp.381-385
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    • 2001
  • Neospora caninum infections have been associated with neonatal paresis as well as abortion around the world. Bovine abortion induced by N caninum was first reported in 1997 in Korea. Diagnosis of N caninum infection is usually based on histopathology and immunohistochemical detection of organism. However, often the tissues having lesion suggestive of N caninum infection were negative on immunohistochemistry. Here, we describe establishment of PCR-based diagnostic strategy for N caninum infection using DNA extracted from paraffin blocks containing the lesion. PCR was able to amplify N caninum-specific bands from the paraffin blocks containing at least moderate degree of inflammation. Compared to paraffin-blocks, DNA extracted from fresh tissues were less sensitive than that of paraffin blocks. This PCR-based method can be practically applicable for rapid diagnosis of bovine N caninum infection with high specificity and sensitivity. Based on this method, 17% of bovine abortion surveyed during a designated period was associated with N caninum infection.

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Hemozoin Pigment: An Important Tool for Low Parasitemic Malarial Diagnosis

  • Mohapatra, Sarita;Ghosh, Arnab;Singh, Ruchi;Singh, Dhirendra Pratap;Sharma, Bhawna;Samantaray, Jyotish Chandra;Deb, Manorama;Gaind, Rajni
    • Parasites, Hosts and Diseases
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    • 제54권4호
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    • pp.393-397
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    • 2016
  • Low parasitemic condition in malaria remains a diagnostic challenge; as the available diagnostic methods failed to detect. Currently, hemozoin (Hz) pigment is gaining attention in the diagnosis of malaria. The major drawback is ease of detection of Hz in routine practice. A pilot study was conducted to evaluate the role of Hz pigment and to compare the performance of quantitative buffy coat assay (QBC) and PCR in such conditions. Clinically suspected cases of malaria were examined by both Giemsa stain and immunochromatographic test (ICT). Samples positive by ICT and negative by Giemsa stain were further examined by nested PCR targeting 18S rRNA and QBC for the presence of malaria parasites and pigments. Thirty blood samples fulfilled the inclusion criteria out of which 23 were Plasmodium vivax (Pv), 4 Plasmodium falciparum (Pf), and 3 mixed (Pv and Pf) by immunochromatographic test. Twenty-one out of 30 (70%) were positive by nested PCR in comparison to 25/30 (83%) by QBC. Samples containing both malaria parasites and Hz pigment by QBC completely showed concordance with the PCR result. However, 61% of total samples containing only Hz pigment were observed positive by PCR. Hz pigment remains an important tool for malaria diagnosis. Identification of leukocytes containing pigments by QBC not only indicates recent malarial infections but also puts light on severity of the disease. QBC assay is a rapid, highly sensitive, and cost-effective method to detect malaria parasites and Hz pigment especially in low parasitemic conditions.

아까바네 바이러스의 분리 및 RT-PCR 진단법에 관한 연구 (Isolation of akabane virus and its molecular diagnosis by reverse transcription polymerase chain reaction)

  • 조재진;이정길;박봉균;장정호;정정원;조인수;안수환
    • 대한수의학회지
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    • 제40권1호
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    • pp.42-48
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    • 2000
  • Akabane disease is transmitted through mosquitoes in cattle, sheep and goats. It shows congenital abnormalities including encephalomyetitis, hydranencephaly, neurogenic arthrogryposis, and deformed neonatal calves. Akabane viruses, 93FMX and K-9 strain, were isolated from fetal matrix of aborted cow and blood of healthy cow, respectively. S gene sequences of 93FMX and K-9 showed 100% homology with that of OBE-1 strain isolated in Japan. Based upon our sequencing data, we synthesized specific primers for PCR diagnosis. Using these primers, we were able to amplify the S gene of Akabane virus not only from the culture fluid of Vero cells but also from the brain tissue of suckling mouse inoculated with, Akabane virus. These PCR products were confirmed by Southern blot hybridization. Not only the sensitivity of PCR test was high enough to detect the viruses of $10^{1.0}TCID_{50}/ml$, but also the time for diagnosis was significantly shorter than that of the virus isolation by tissue culture method. This method was also effective for the detection of Akabane virus in the cerebrum of fetus. RT-PCR method may be used for a useful diagnostic test of the clinical cases of Akabane disease.

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Ultra Real-Time PCR을 활용한 Avian Influenza Virus Subtype의 조기진단법 (Early Diagnostic Method of Avian Influenza Virus Subtype Using Ultra Real-Time PCR)

  • 김상태;김영균;김장수
    • 미생물학회지
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    • 제47권1호
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    • pp.30-37
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    • 2011
  • 조류 인플루엔자 바이러스(AIV) 아형을 ultra-time PCR법(UPCR)을 이용하여 초스피드로 진단할 수 있는 방법을 고안하였다. 표적 대상의 프라이머는 AIV H5N1 아형의 hemagglutinin(HA) 유전자 중 가장 상보성이 높은 133 bp의 부위를 선택하였고, 실험의 안전을 위하여 인공합성의 방법으로 제작하였다. 압타머와 결합한 molecular beacon 기반 Mini-Opticon Q-PCR 기기를 사용한 UPCR법으로, 총 UPCR 반응액의 양을 10 ${\mu}l$으로, UPCR과 용융온도 분석시간을 15분 이내로 매우 짧게 단축시켰다. 민감도 측정에서 최소의 주형인 5분자의 HA 유전자만으로 정확히 AIV의 특이적 133 bp를 합성하였다. UPCR로 디자인된 이 PCR은 AIV 아형의 진단에 적용될 수 있을 뿐 아니라, UPCR이 기반되는 진단을 이용하여 다른 병원체에도 널리 적용 될 수 있을 것으로 기대된다.

PCR for Diagnosis of Male Trichomonas vaginalis Infection with Chronic Prostatitis and Urethritis

  • Lee, Jong-Jin;Moon, Hong-Sang;Lee, Tchun-Yong;Hwang, Hwan-Sik;Ahn, Myoung-Hee;Ryu, Jae-Sook
    • Parasites, Hosts and Diseases
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    • 제50권2호
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    • pp.157-159
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    • 2012
  • The aim of this study was to assess the usefulness of PCR for diagnosis of Trichomonas vaginalis infection among male patients with chronic recurrent prostatitis and urethritis. Between June 2001 and December 2003, a total of 33 patients visited the Department of Urology, Hanyang University Guri Hospital and were examined for T. vaginalis infection by PCR and culture in TYM medium. For the PCR, we used primers based on a repetitive sequence cloned from T. vaginalis (TV-E650). Voided bladder urine (VB1 and VB3) was sampled from 33 men with symptoms of lower urinary tract infection (urethral charge, residual urine sensation, and frequency). Culture failed to detect any T. vaginalis infection whereas PCR identified 7 cases of trichomoniasis (21.2%). Five of the 7 cases had been diagnosed with prostatitis and 2 with urethritis. PCR for the 5 prostatitis cases yielded a positive 330 bp band from bothVB1 and VB3, whereas positive results were only obtained from VB1 for the 2 urethritis patients. We showed that the PCR method could detect T. vaginalis when there was only 1 T. vaginalis cell per PCR mixture. Our results strongly support the usefulness of PCR on urine samples for detecting T. vaginalis in chronic prostatitis and urethritis patients.

다중 중합효소 연쇄반응을 이용한 소의 Johne병 진단 기법 확립 (Diagnosis of Bovine Johne's Disease Using Multiplex Polymerase Chain Reactions)

  • 김종배;송혜원;김근희;김홍;신광순;김두
    • 대한의생명과학회지
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    • 제6권1호
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    • pp.65-72
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    • 2000
  • 반추수에서 발생하는 Johne병의 조기 진단 방법을 제시하고 이 질병의 원인체와 미생물학적 특징 이 유사한 M. bovis, M. avium 등의 mycobacteria 감염증을 감별 진단하는 방법 을 개발하기 위하여 Mycobacterium 균속의 표준균주를 사용하여 중합효소 연쇄반응을 확립하였다. Johne병으로 의심되는 소의 혈액과 유즙을 채취하여 분리한 단핵구 및 거식세포로부터 genomic DNA를 추출하였다. 각 시료로부터 추출한 DNA를 template로 이용하여 Mycobacterium spp.에 특이적인 16S rDNA primer set를 이용한 PCR을 수행하여 시료내의 mycobacterial DNA 보유 여부를 확인하였다. 한편 mycobacteria 양성으로 확인된 시료는 M. avium complex 균종에 특이한 16S rDNA 염기서열을 기초로 하여 제작한 primer set와 M. paratuberculosis의 IS900 sequence에 특이한 primer set를 이용하여 duplex PCR을 수행하여 Johne병 원인체의 보균 여부를 조사하였으며, 이 결과를 oligonucleotide probe를 사용한 Southern blot hybridization을 통하여 다시 확인하였다. 이와 같은 duplex PCR기법을 실제 축산 현장에서 수집한 유즙과 말초혈액으로부터 분리한 단핵구 및 거식세포 시료에 적용한 결과 본 연구에서 확립한 duplex PCR기법 유용성을 확인할 수 있었다.

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