• Korean Journal of Clinical Laboratory Science
• /
• v.47 no.4
• /
• pp.313-317
• /
• 2015

Due to the increase in incidence of infection of Mycobacterium tuberculosis complex (MTC), it is imperative that a rapid diagnosis accompanies the handling of MTC. This is due to the three to eight weeks it takes to culture Mycobacteria, and the lack of sensitivity of microscopic examination of AFB. Recently, nested PCR has been used to detect and diagnose mycobacteria. It is especially useful in complementing diagnosis by histological extra pulmonary. After culturing all the specimens and practicing the nested PCR, we did comparison analysis between nested PCR and culture. There were 76 specimens, 31 of which were positive. Of the 31 positive specimens in culturing, only 22 were positive in nested PCR. Of the 45 negative specimens, 36 were negative in nested PCR. As a result, Sensitivity was 71% and specificity was 80%. Furthermore, the positive predictive value was 71% and negative predictive value was 80%. These results indicate that nested PCR based techniques are sensitive, specific, and rapid methods for the detection of MTC.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.6
• /
• pp.1164-1169
• /
• 2008

We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) and 9 neuraminidase (N1-N9) subtypes that are commonly isolated from avian, swine, and human influenza A viruses. RT-PCR products with unique sizes characteristic of each subtype were amplified by multiplex RT-PCRs, and sequence analysis of each amplicon was demonstrated to be specific for each subtype with 24 reference viruses. The specificity was demonstrated further with DNA or cDNA templates from 7 viruses, 5 bacteria, and 50 influenza A virus-negative specimens. Furthermore, the assays could detect and subtype up to $10^5$ dilution of each of the reference viruses that had an original infectivity titer of $10^6\;EID_{50}/ml$. Of 188 virus isolates, the multiplex RT-PCR results agreed completely with individual RT-PCR subtyping results and with results obtained from virus isolations. Furthermore, the multiplex RT-PCR methods efficiently detected mixed infections with at least two different subtypes of influenza viruses in one host. Therefore, these methods could facilitate rapid and accurate subtyping of influenza A viruses directly from field specimens.

• Journal of Korean Academy of Fundamentals of Nursing
• /
• v.6 no.3
• /
• pp.359-368
• /
• 1999

Background : The purpose of this study was to determine whether cleansing the perineum and urethral meatus and using midstream urine affect the rate of bacterial contamination of urine specimens, and to determine the optimum urine collection method. We studied 41 asymptomatic healthy nursing school students. Women who were menstruating were not excluded from this study. Method : The first and midstream urine samples were collected during consecutive urinationsby each woman. The first sample was not a clean-catch specimen, and the second one was a clean-catch specimen. Both specimens were studied by urinalysis and bacterial culture with standard methods. Results : 41 women met the study criteria and 39 successfully completed the study. None of the urine cultures were positive. 68.3% of the non clean-catch first urine cultures, 53.7% of the non clean-catch midstream cultures, 33.3% of the first clean-catch urine culteres and 30.8% of the midstream clean-catch urine were found to be contaminated. There was a significant difference in the bacterial contamination rates between the first and midstream urine, and the clean-catch and non clean-catch urine(p=0.035, p =0.001 respectively). On urinalysis, 7.3% of the non clean-catch first urine, 7.3% of the non clean-catch midstream urine, 2.6% of the clean-catch first urine and 2.6% of clean-catch midstream urine were found to be above grade 2. Conclusions : According to our results, the bacterial contamination rate was the lowest in midstream and clean catch urine specimens. Threrfore it is recommended that the midstream clean-catch technique is the standard practice for collecting urine specimens for bacterial culture in women.

• Journal of Yeungnam Medical Science
• /
• v.32 no.2
• /
• pp.71-79
• /
• 2015

Background: Recent studies have shown that the nontuberculosis mycobacterium (NTM) recovery rate in clinical cultures has increased within Korea. However, another study conducted by a secondary hospital within Daegu reported different results. Therefore, the purpose of this study is to understand and evaluate the microbiological distribution and clinical features of NTM in Daegu. Methods: A retrospective study was conducted on 11,672 respiratory specimens undergoing acid fast bacilli (AFB) culture from 6,685 subjects who visited Yeungnam University Respiratory Center from January 2012 to December 2013. Results: Of the 11,672 specimens undergoing AFB culture, 1,310 specimens (11.2%) showed positive results. Of these specimens, NTM was recovered from 587 specimens, showing a recovery rate of 44.8%. Identification test for NTM was performed on 191 subjects; the results were as follows: M. avium-intracellulare complex (MAC) 123 (64.4%), M. abscessus 20 (10.5%), M. kansasii 12 (6.3%), and 33 other NTM germ strains. Of the 382 subjects with NTM, 167 were diagnosed with pulmonary NTM disease (43.7%), however virulence differed depending on NTM strain. Multivariate analysis showed that nodular bronchiectasis, the nodules, and finding consistent with cavity under imaging study were statistically significant for triggering pulmonary NTM disease. AFB culture showing MAC and M. abscessus was statistically significant as well. Positive predictive value for NTM polymerase chain reaction (NTM-PCR) was 88.6%. Conclusion: Results for NTM recovery rate within the Daegu area were similar to those for the Seoul metropolitan area. We can assume that NTM infection is increasing in our community, therefore AFB-positive subjects (1) should undergo NTM-PCR, (2) should have their culture results checked for differentiation of mycobacterium tuberculosis complex (MTB) from NTM, and (3) undergo NTM identification test to confirm its type. Administration of treatment with the above results should be helpful in improving the patients' prognosis.

• Korean Journal of Clinical Laboratory Science
• /
• v.36 no.2
• /
• pp.76-88
• /
• 2004

The purpose of this study was to investigate the distribution of Enterococci isolated from clinical specimens, and identify the aspect of antibiotic susceptibility and analyze the genetic difference by executing Rep-PCR over the strains resistant to aminoglycoside-typed antibiotics. From an assortment of the clinical specimens, 100 strains were isolated. The collection consisted of 49 strains of E. faecalis, 34 strains of E. faecium, 9 strains of E. avium, 4 strains of E. gallinarum, 3 strains of E. casseliflavus, and 1 strain of E. hirae. Ninety five were isolated from inpatients, and five strains were isolated from outpatient. Most of the E. faecalis and E. faecium were originated from urine, pus, and sputum. Most Enterococci showed 80% resistance to the cephalosprin-typed antibiotics. E. faecium showed the high resistance to all the antibiotic substances. One tenths of Enterococci showed the resistance to vancomycin. And also, most Enterococci showed the high resistance to amikacin and gentamicin as aminoglycoside-typed antibiotics. Genetic diversity of the resistant strains to aminoglycoside estimated using Rep-PCR was not significanty different.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.3
• /
• pp.953-956
• /
• 2016

The risk of developing cervical cancer in women infected with human papillomavirus (HPV) may be influenced by an individual's genetic susceptibility. Published data linking single nucleotide polymorphisms (SNPs) in the tumor necrosis factor-alpha (TNF-${\alpha}$) promoter region at positions -308G>A (rs1800629) and -238G>A (rs361525) to cervical cancer risk have been inconclusive. In this study, we examined 251 cervical specimens and classified them into two groups according to their cytological findings: 121 cancer cases and 130 controls (low-grade squamous intraepithelial lesion and normal cytology). All specimens were typed by PCR and sequencing for TNF-${\alpha}$ promoter -308G>A (rs1800629) and -238G>A (rs361525). The genotype distribution of SNPs in either rs1800629 or rs361525 did not significantly demonstrate higher frequency in the cancer group (p=0.621 and p=0.68, respectively). Based on these results, neither the TNF-${\alpha}$ promoter -308G>A (rs1800629) nor the -238G>A (rs361525) polymorphism presents a major risk factor for cervical cancer among Thai women. Larger studies are necessary to elucidate possible genetic mechanisms influencing cervical cancer development.

• Korean Journal of Clinical Laboratory Science
• /
• v.39 no.1
• /
• pp.7-18
• /
• 2007

A total of 1,780 isolates of Enterococcus spp. were isolated from 63,133 clinical specimens from Dec 1, 2005 to Nov 1, 2006 in "C" hospital. Isolation frequencies of Enterococcus spp. were 50.9% for E. faecalis, 41.7% for E. faecium, and 7.4% for other Enterococcus spp. containing E. avium, E. gallinarum, E. casseliflavus, E. durans, E. hirae, and E. raffinosus. There were no significant difference between gender, but according to the age group analysis, Enterococcus spp. were more frequently isolated in patients over 50 years old (20.0~24.6%) than those isolated from the patients under the age of 0~49 (1.3~9.4%). In monthly analysis, Enterococcus spp. were the most frequently isolated in April (11.9%), but presented at lowest frequency in February (5.2%). Seasonal analysis did not show a significant difference. Over half of enterococci were isolated from random urine (44.9%) and catherterized urine (15.7%). Frequencies of vancomycin resistant E. faecalis and E. faecium were 0.1% and 31.0%, respectively. Teicoplanin resistant Enterococcus was 13.3% in E. faecalis, 17.6 % in E. faecium. The Enterococcus species showing over 80% susceptibility against antimicrobial agents were E. faecalis, E. durans and E. hirae in vancomycin; E. faecalis, E. gallinarum, E. casseliflavus, E. durans and E. hirae in ampicillin. The antimicrobial agent showing susceptibility against whole group of Enterococcus species was only linezolid (95.9%), and a selection of antimicrobial agent is necessary to do essential performance identification and susceptibility tests.

• Korean Journal of Clinical Laboratory Science
• /
• v.43 no.1
• /
• pp.1-5
• /
• 2011

The infection rate of syphilis is still increasing in the world especially in developing countries and the infection is often seen in large amounts of clinical specimens. For the diagnosis of this disease, Rapid Plasma Reagin (RPR)/Venereal Disease Research Laboratory (VDRL) has still been used as one of major primary methods to diagnose syphilis even though the test readings are somewhat subjective with high false positive rates. Recently, the automatic ARCHITECT Syphilis TP, which is based on the detection of the TP-specific antibodies, has been introduced in many laboratories. Therefore, the clinical assessment of the method is needed to provide primary diagnosis of syphilis at the moment. We evaluated 3 different manual rapid kits and ARCHITECT Syphilis TP comparing with RPR/FTA-ABS and analysed their diagnostic properties. From February 2006 to April 2008, 203 positive and 250 negative specimens, obtained from Chungbuk National University Hospital were used for the evaluation. In the evaluation between manual rapid kits, their specificities were as high as 99.2 ~ 99.6% while their sensitivities were observed with little differences; 98.0% (199/203) for Kit A, 96.6% (196/203) for Kit B, and 97.4% (197/203) for Kit S. In the case of ARCHITECT Syphilis TP test, it showed 100% specificity (250/250) and 98.5% sensitivity (249/250). Kappa values comparing with RPR/FTA-ABS were 0.978 for Kit A, 0.964 for Kit B and Kit S, and 0.987 for ARCHITECT Syphilis TP. From our evaluation, we found out that manual rapid tests and ARCHITECT Syphilis TP have very good clinical accuracies and high kappa agreements with RPR/FTA-ABS. Due to its automation and quick simultaneous diagnosis with another serological markers, we suggest that the ARCHITECT Syphilis TP is one of best suitable method for the primary diagnosis of syphilis and that it might be able to replace RPR method in the laboratories.

• Korean Journal of Clinical Laboratory Science
• /
• v.46 no.3
• /
• pp.99-105
• /
• 2014

Isolating total DNA from small samples using traditional methods is difficult and inefficient mainly due to loss of DNA during filtration and precipitation. With advances in molecular pathology, DNA extraction from micro-dissected cells has become essential in handling clinical samples. Genomic DNA extraction using small numbers of cells can be very important to successfully PCR amplify DNA from small biopsy specimens. We compared our experimental genomic DNA extraction method (A) with two other commercially available methods: using spin columns (B), and conventional resins (C), and determined the efficacy of DNA extraction from small numbers of cells smeared on a glass slide. Approximately 50, 100, 200, 500 and 1000 cells were isolated from fine needle aspiration biopsy (FNAB) slides aspirated from histologically proven papillary thyroid carcinoma masses. DNA was extracted using the three techniques. After measuring DNA quantity, PCR amplification was performed to detect the ${\beta}$-globin and $BRAF^{V600E}$ gene mutations. DNA extracted by method (A) showed better yield than the other methods in all cell groups. With our method, a suitable amount of genomic DNA to produce amplification was extracted from as few as 50 cells, while more than 100 to 200 cells were required when methods (B) or (C) were applied. Our genomic DNA extraction method provides high quality and improved yields for molecular analysis. It will be especially useful for paucicellular clinical samples which molecular pathologists often confront when handling fine needle aspiration cytology, exfoliative cytology and small biopsy specimens.

• Anatomy and Cell Biology
• /
• v.56 no.2
• /
• pp.200-204
• /
• 2023

Although in counterpart, the sphenopalatine artery (SPA), has been well described in the medical literature, the sphenopalatine vein (SPV) has received scant attention. Therefore, the present anatomical study was performed. Additionally, we discuss the variations, embryology, and clinical significance of the SPV. Adult cadaveric specimens underwent dissection of the SPV. In addition, some specimens were submitted for histological analysis of this structure. The SPV was found to drain from the sphenoidal sinus and nasal septum. Small tributaries traveled through the nasal septum with the posterior septal branches of the SPA and nasopalatine nerve. The SPA and SPV were found to travel through the sphenopalatine foramen and another tributary was found to perforate the medial plate of the pterygoid process and to connect to the pterygoid venous plexus which traveled lateral to the medial plate of the pterygoid process. The vein traveled through the posterior part of the lateral wall of the nasal cavity with the posterior lateral nasal branches of the SPA and the lateral superior posterior nasal branches of the maxillary nerve. To our knowledge, this is the first anatomical study on the SPV in humans. Data on the SPV provides an improved anatomical understanding of the vascular network of the nasal cavity. Developing a more complete picture of the nasal cavity and its venous supply might help surgeons and clinicians better manage clinical entities such as posterior epistaxis, cavernous sinus infections, and perform endoscopic surgery with fewer complications.