Detection of protozoan parasites Perkinsus sp. and P. atlanticus was developed in this study using a specific polymerase chain reaction (PCR) to diagnose the presence of those organisms that causes extensive mortalities of marine shellfishes. The PCR was conducted together with fluid thioglycollate medium (FTM) method and 2 M NaOH lysis method. For the test, Manila clams, Ruditapes philippinarum, were collected from four coastal locations in Korea including Wando Island, Gimnyeong, Sungsan and Sogwipo in Jeju. In addition, trophozites of Perkinsus sp. cultivated in vitro and the granular ark clam, Tegillarca granosa, taken from Gangjin on the south coast of Korea, were used as positive and negative controls, respectively. Expected DNA bands were detected in the samples from Wando Island, Sungsan and the in vitro cultured Perkinsus sp. when the probes specific for the genus Perkinsus and P. atlanticus were used. The samples were also positively diagnosed by the FTM and 2 M NaOH methods. In contrast, the Manila clams from Gimnyeong and Sogwipo, and the granular arks clams from Gangjin showed no detectable signs of infection with the PCR, the FTM method and the 2 M NaOH lysis method. On the other hand, being amplified by p. atlanticus specific primer, it is suggested that the protozoan parasite Perkinsus sp. found in the Korean Manila clam is P. atlanticus. Finally the PCR- based assay developed in the present study can be used in detection of Perkinsus infection and discrimination of Peykinsus species in quarantine stations or laboratories due to the high sensitivity and specificity as well as its rapid detection.
Kim, Yeun;Park, Sunyoung;Kim, Jungho;Chang, Yunhee;Ha, Sunmok;Choi, Yeonim;Lee, Hyeyoung
Korean Journal of Clinical Laboratory Science
/
v.50
no.1
/
pp.20-28
/
2018
Tuberculosis (TB) is a bacterial infection disease caused by members of the species Mycobacterium tuberculosis (MTB) complex. Approximately one third of the world's population is infected with TB. In Korea, approximately 40,000 new patients are identified each year. Moreover, infections from non-tuberculous mycobacteria (NTM) have also increased. In the diagnosis of TB and NTM, traditional bacterial cultures are required for 3 to 4 weeks. Therefore, rapid and accurate diagnostic tests for TB and NTM are needed. To distinguish between TB and NTM, a range of diagnostic methods have been developed worldwide. In vitro diagnostic assays are constantly being developed to meet the increasing need for the rapid and accurate identification for TB and NTM. On the other hand, the performance evaluations of in vitro diagnostic reagents for TB and NTM are lacking. Recently, the Korea Food and Drug Administration (KFDA) issued a guideline for in vitro diagnostic reagents for MTB and NTM. Here, this study analyzed the performance of currently developed in vitro diagnostic reagents for TB and NTM in the US FDA. This analysis of US FDA approved molecular assays could serve as a useful reference for an evaluation of the reagent performance of TB and NTM.
Kim, Jin-Eui;Kim, Jung-Soo;Choi, Nam-Gil;Han, Jae-Bok
The Journal of the Korea Contents Association
/
v.17
no.11
/
pp.50-59
/
2017
To evaluate the capability of integrated PET-MRI, which has recently been utilized in the clinical practices, on the diagnosis of liver cancer, its utility was assessed by $2{\times}2$ decision matrix. The numbers of abnormal and normal decisions on the liver cancer were 98 and 51 cases, respectively, upon PET-MRI scan results of the subjects, and the numbers of positive and negative decisions were 103 and 62, respectively, upon cytopathologic results. Out of the two tests, 95 cases were shown as true-positive and 3 were false positive, while 62 were true negative and 5 were false negative. Upon the results of PET-MRI test, its sensitivity, specificity, false negative rate, and false positive rate were 95.00%, 95.38%, 0.05%, and 95.15%, respectively. Therefore, it is considered to have the high potential to use the determination of the stage before the surgery, detections of recurrence and remote metastasis, assessment of uncertain remote lymph node metastasis, and so on in the diagnosis of the liver cancer, and also for the clinical utility of PET-MRI to be sufficient by integrated diagnosis and follow up scan with pathological studies.
Objective: The serum S100 protein has been known to reflect the severity of neuronal damage. The purpose of this study was to assess the prognostic value of the serum S100 protein by Elecsys S100 immunoassay in patients with subarachnoid hemorrhage (SAH) and intracerebral hemorrhage (ICH) and to establish reference value for this new method. Methods: Serum S100 protein value was measured at admission, day 3 and 7 after bleeding in 42 consecutive patients (SAH : 20, ICH : 22) and 74 healthy controls, prospectively. Admission Glasgow coma scale (GCS) score, Hunt & Hess grade and Fisher grade for SAH, presence of intraventricular hemorrhage, ICH volume, and outcome at discharge were evaluated. Degrees of serum S100 elevation and their effect on outcomes were compared between two groups. Results: Median S100 levels in SAH and ICH groups were elevated at admission (0.092 versus $0.283{\mu}g/L$) and at day 3 (0.110 versus $0.099{\mu}g/L$) compared to healthy controls ($0.05{\mu}g/L;$ p<0001). At day 7, however, these levels were normalized in both groups. Time course of S100 level in SAH patient was relatively steady at least during the first 3 days, whereas in ICH patient it showed abrupt S100 surge on admission and then decreased rapidly during the next 7 days, suggesting severe brain damage at the time of bleeding. In ICH patient, S100 level on admission correlated well with GCS score (r=-0.859; p=0.0001) and ICH volume (r=0.663; p=0.001). A baseline S100 level more than $0.199{\mu}g/L$ predicted poor outcome with 92% sensitivity and 90% specificity. Logistic regression analyses showed Ln (S100) on admission as the only independent predictor of poor outcome (odd ratio 36.1; 95% CI, 1.98 to 656.3) Conclusion: Brain damage in ICH patient seems to develop immediately after bleeding, whereas in SAH patients it seems to be sustained for few days. Degree of brain damage is more severe in ICH compared to SAH group based on the S100 level. S100 level is considered an independent predictor of poor outcome in patient with spontaneous ICH, but not in SAH. Further study with large population is required to confirm this result.
Kim, Seung Hyun;Choi, Jun Ho;Hwang, Jae Ha;Kim, Kwang Seog;Lee, Sam Yong
Archives of Craniofacial Surgery
/
v.21
no.5
/
pp.276-282
/
2020
Background: Orbital fractures are the most common pediatric facial fractures. Treatment is conservative due to the anatomical differences that make children more resilient to severe displacement or orbital volume change than adults. Although rarely, extensive fractures may result in enophthalmos, causing cosmetic problems. We aimed to establish criteria for extensive fractures that may result in enophthalmos. Methods: We retrospectively reviewed the charts of patients aged 0-15 years diagnosed with orbital fractures in our hospital from January 2010 to February 2019. Computed tomography images were used to classify the fractures into linear, trapdoor, and open-door types, and to estimate the defect size. Data on enophthalmos severity (Hertel exophthalmometry results) and fracture pattern and size at the time of injury were obtained from patients who did not undergo surgery during the follow-up and were used to identify the surgical indications for pediatric orbital fractures. Results: A total of 305 pediatric patients with pure orbital fractures were included-257 males (84.3%), 48 females (15.7%); mean age, 12.01±2.99 years. The defect size (p=0.002) and fracture type (p=0.017) were identified as the variables affecting the enophthalmometric difference between the eyes of non-operated patients. In the linear regression analysis, the variable affecting the fracture size was open-door type fracture (p<0.001). Pearson's correlation analysis demonstrated a positive correlation between the enophthalmometric difference and the bony defect size (p=0.003). Using receiver operating characteristic curve analysis, a cutoff value of 1.81 ㎠ was obtained (sensitivity, 0.543; specificity, 0.724; p=0.002). Conclusion: The incidence of enophthalmos in pediatric pure orbital fractures was found to increase with fracture size, with an even higher incidence when open-door type fracture was a cofactor. In clinical settings, pediatric orbital fractures larger than 1.81 ㎠ may be considered as extensive fractures that can result in enophthalmos and consequent cosmetic problems.
Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.
Proceedings of the Korean Society of Toxicology Conference
/
2006.11a
/
pp.46-54
/
2006
The methylmercury (MeHg) is a toxic environmental pollutant, causing serious neurological and developmental effects in humans. Recent epidemiological studies have indicated that ingestion of MeHg in fish during pregnancy can result in neuroethological effects in the offspring. However, the mechanism underlying the MeHg-toxicity is not fully understood. To elucidate the mechanisms of toxicity of MeHg and of defense against MeHg, we searched for factors that determine the sensitivity of yeast cells to MeHg, and found that overexpression of Cdc34, a ubiquitin-conjugating enzyme (E2) that is a component of the ubiquitin-proteasome (UP) system, induces a resistance to MeHg toxicity in both yeast and human cells. The UP system is involved in the intracellular degradation of proteins. When Cdc34 is overexpressed in cells, ubiquitination reactions are activated and the degradation of certain proteins by the UP system is enhanced. Therefore, it seems likely that certain as-yet-unidentified proteins that increase MeHg toxicity might exist in cons and that toxicity might be reduced by the enhanced degradation of such proteins, mediated by the UP system, when Cdc34 is overexpressed. SCF ubiquitin-ligase is a component of UP system and consists of Skpl, the scaffold protein Cdc53, the RING-finger protein Hrt1, and one member of the family of F-box proteins. The F-box proteins directly bind to the substrates and are the determinants of substrate specificity of SCF. Therefore, we searched for the f-box protein that cofers resistance to MeHg, and found that overexpression of Hrt3 or Yi1224w induced resistance to MeHg toxicity in yeast cells. Since the protein(5) that enhance toxicity of MeHg might plausibly be induced in substrates of both f-box proteins, we next searched for substrate proteins that are recognized by Hrt3 or Y1r224w using two-hybrid screen. We found that Did3 or Crsl interacts with Hrt3; and Eno2 interacts with Yir224w. The yeast cells that overexpressed each those proteins showed hypersensitivity to MeHg, respectively, indicating that those proteins enhance the MeHg toxicity. Both Dld3 and Eno2 are proteins involved in the synthesis of pyruvate, and overexpression of both proteins might induce increase in interacellular levels of pyruvate. Deletion of Yi1006w that transports pyruvate into the mitochondria induced aresistance to MeHg. These results suggest that the promotion of the pyruvate irdlowinto the mitochondria might enhance MeHg toxicity. This study providesimportant keyfor the elucidauon of the molecular mechanism of MeHg toxicity.
In the study, we developed and evaluated a uracil N-glycosylase (UNG)-supplemented single-tube nested reverse transcription-polymerase chain reaction (UsnRT-PCR) assay that can carried out first-round RT-PCR and second-round nested PCR in a reaction tube without reaction tube opening and can simultaneously detect EU- and NA-PRRSV. The UsnRT-PCR confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PRRSV DNA from previous UsnRT-PCR. Primer specificities were evaluated with RNAs extracted from 8 viral strains and our results revealed that the primers had a high specificity for both genotypes of PRRSV. The sensitivity of the UsnRT-PCR was 0.1 $TCID_{50}$/0.1 mL for EU- or NA-PRRSV, respectively, which is comparable to that of previously reported real time RT-PCR (RRT-PCR). Clinical evaluation on 110 field samples (60 sera and 50 lung tissues) by the UsnRT-PCR and the RRT-PCR showed that detection rates of the UsnRT-PCR was 70% (77/110), and was relatively higher than that of the RRT-PCR (69.1%, 76/110). The percent positive or negative agreement of the UsnRT-PCR compared to RRT-PCR was 96.1% (73/76) or 90.9% (30/33), showing that the test results of both assays may be different for some clinical samples. Therefore, it is recommend that diagnostic laboratory workers use the two diagnostic assays for the correct diagnosis for the relevant samples in the swine disease diagnostic laboratories. In conclusion, the UsnRT-PCR assay can be applied for the rapid, and reliable diagnosis of PRRSV without concerns about preamplified DNA carryover contamination that can occurred in PCR process in the swine disease diagnostic laboratories.
The purpose of this study was to evaluate the clinical usefulness of I-123 MIBG scintigraphy with early planar and SPECT image in the diagnosis of neuroendocrine tumors. We reviewed I-123 MIBG scintigraphies of 21 patients who had been suspected to have neuroendocrine tumors by CT or MRI findings. Early 4 hour planar and SPECT images were obtained in all patients and delayed (13-24 hour) planar images were performed in 17 patients. Final diagnoses were made by surgery, biopsy, or clinical follow up. Twelve patients were confirmed to have neuroendocrine tumors. With 4 hour planar and SPECT images, there were 9 true positives(6 pheochromocytomas, 1 paraganglioma, 1 neuroblastoma, and 1 medullary cancer of the thyroid), 8 true negatives(1 adrenal cortical adenoma, 1 malignant fibrous histiocytoma, 1 adenoma in colon and 5 benign nonfunctioning adrenal tumors), 1 false positive(hepatocellular carcinoma) and 3 false negatives(1 recurred medullary cancer of the thyroid, 1 liver metastasis of carcinoid tumor and 1 ganglioneuroma). The sensitivity and specificity of I-123 MIBG scintigraphy were 75% and 89%, respectively. SPECT images provided good anatomical correlation with CT or MRI. Delayed images showed increased tumor to background ratio in 5 out of 8 true positive patients, but did not change the diagnosis. In conclusion, early 4 hour images with I-123 MIBG is clinically convenient and useful method in the detection of neuroendocrine tumors, and SPECT images can provide good anatomical correlation with CT or MRI.
Detection of protozoan parasites Perkinsus sp. and P. atlanticus was developed in this study using a specific polymerase chain reaction (PCR) to diagnose the presence of those organisms that causes extensive mortalities of marine shellfishes. The PCR was conducted together with fluid thioglycollate medium (FTM) method and 2 M NaOH lysis method. For the test, Manila clams, Ruditapes philippinarum, were collected from four coastal locations in Korea including Wando Island, Gimnyeong, Sungsan and Sogwipo in Jeju. In addition, trophozites of Perkinsus sp. cultivated in vitro and the granular ark clam, Tegillarca granosa, taken from Gangjin on the south coast of Korea, were used as positive and negative controls, respectively. Expected DNA bands were detected in the samples from Wando Island, Sungsan and the in vitro cultured Perkinsus sp. when the probes specific for the genus Perkinsus and P. atlanticus were used. The samples were also positively diagnosed by the FTM and 2 M NaOH methods. In contrast, the Manila clams from Gimnyeong and Sogwipo, and the granular arks clams from Gangjin showed no detectable signs of infection with the PCR, the FTM method and the 2 M NaOH lysis method. On the other hand, being amplified by p. atlanticus specific primer, it is suggested that the protozoan parasite Perkinsus sp. found in the Korean Manila clam is P. atlanticus. Finally the PCR- based assay developed in the present study can be used in detection of Perkinsus infection and discrimination of Peykinsus species in quarantine stations or laboratories due to the high sensitivity and specificity as well as its rapid detection.
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