The quality control of pathological specimens is important for accurate molecular pathology testing. This study evaluated that specimen factors affecting the DNA quality during tissue processing and sample types for BRAF, EGFR, and KRAS mutations tests. One thousand seven hundred and seventy-two molecular pathology tests were investigated for the factors influencing the DNA quality, such as sample type, formalin fixation time, and reexamination status. Cytology samples stored in a saline solution had better DNA quality than commercial cytology preservation. Tissue samples fixed in formalin within 24 hours had better DNA quality than the samples fixed over 24 hours. Between the types of samples, fresh tissue samples and tissue samples with a high tumor cell density had relatively better DNA quality than the formalin-fixed paraffin-embedded (FFPE) tissues and cytology specimens. Of real-time PCR, the non-PNA Ct value increased proportionally with samples held for longer than 24 hours in formalin, and that the formalin-fixed time affects the sample DNA quality. In conclusion, the appropriate tumor cellularity and 10% neutral formalin fixation time are the most important factors for maintaining the DNA quality. These factors should be managed properly for an accurate pathological molecular test to ensure optimal DNA quality.
Kim, Jung In;Kang, Mi Ji;Kim, Na Kyung;Park, Ji Sol;Kwon, Won Hyun;Lee, Kyung Jae
The Korean Journal of Nuclear Medicine Technology
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v.25
no.2
/
pp.29-34
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2021
Purpose Sample reception environment system in nuclear medicine has not changed much compared to 20 years ago. When preparing sample for in vitro test, there was no significant change because the test was carried out by generating an own specimen from the parent specimen. In this study, We would like to introduce a method that automatically removes the sample cap using the automated decapper equipment and enables automatic reception at the same time. In addition, including a provisional reception system. Materials and Methods In 2019, it was intended to get a device that automatically removes the cap of a patient's blood sample. This equipment is the same as the equipment used in the Department of Laboratory Medicine (Vacuette Ⓡ Unicap Belt Decapper, Greiner bio-one, Austria). However, the purchase was delayed due to differences in tube size, budget, and space. In January 2020, we borrowed domestic automatic decapper equipment and modified it to suit our laboratory environment. After 9 months, we were able to introduce a system that automatically removes the lid of a patient's blood sample and at the same time automatically accepts the test. And, through the provisional reception system, it was possible to know the arrival of the specimen in a short time. Results With the use of an automatic decapper device, the sample cap was automatically removed, and the reception proceeded at the same time. So, it was very efficient at work because it shortened the sample preparation time by about 20 minutes. In addition, it was possible to prevent the examiner's musculoskeletal disorders caused by repeated wrist use. After using the provisional reception system, patients were able to be discharged quickly, and the number of phone calls to confirm the arrival of samples was reduced. Conclusion Most hospitals have about four employees in the nuclear medicine in vitro laboratory. It is effective to use automatic decapper equipment and a provisional reception system for organizations that perform work with the minimum number of personnel.
Purpose: Autopipetting system is an efficient automated equipment pipetting patient samples and reagents for rapid and accurate test. However, it can cause carryover between high concentration sample and low concentration sample. We evaluated carryover contamination of TECAN freedom Evo 100 autopipetting system. Materials and Method: We studied carryover contamination of $\alpha$-fetoprotein (AFP) and carcinoembryonic antigen (CEA) test on TECAN freedom Evo 100 autopipetting system. Very low concentration control samples were pipetted for comparison to the contaminated very low concentration samples. Then, The contaminated very low concentration samples were pipetted following the high concentration samples were pipetted alternately. The difference of low concentration samples represents carryover. The target value to decide carryover was 1ppm (parts per million). Results: For AFP, the mean values of the uncontaminated control samples and the contaminated samples were less than 0.6 IU/mL (the l imit of detection (LoD)). Carryover did not occur even though the high concentration sample which value was 650000 IU/mL. For CEA, the values of the low concentration control samples and the contaminated samples were less than 0.2 ng/mL (LoD). Carryover did not occur even though the high concentration sample which value was 65,000 ng/mL. Conclusions: Sample carryover was not found on TECAN freedom Evo 100 autopipetting system for AFP, CEA. However, carryover is a potential problem with automated instruments and robotic pipetting systems. Therefore, Clinical laboratories must periodically verify carryover contamination for the accurate and confidential test results.
Journal of Korean Society of Environmental Engineers
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v.32
no.10
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pp.979-985
/
2010
This study was performed to detect enterococci strain as an indicator of faecal contamination, to identify of 16S rDNA sequence and vancomycin resistance by MIC (Minimum Inhibitory Concentration) test from drinking spring-water samples in Seoul. The detection frequency of enterococci was 42 (19.8%) among 212 samples, and its concentration was ranged from 0 to 110 CFU/100 mL. These results were confirmed the possibility as an indicator microorganisms that similar to the frequency of E. coli detection (t test p-value 0.268, significant level 0.05). Isolated 56 enterococci samples were identified by 16S rDNA sequence data and their NCBI BLAST searching. They were identified to Enterococcus faecalis of 24 samples, E. faecium (10), E. casseliflavus (10), E. gallinarum (3), E. hirae (2), E. durans (2), E. sanguinicola (1). E. faecalis was dominant species that clinical case report of a domestic was similar. Vancomycin resistant enterococci (VRE) of 53 samples showed that vanB and vanC1/C2 type with 2 and 12 case, respectively. These results indicated that the drinking spring-water quarantined to fecal pollution for block out outbreak of gastrointestinal symptom with using such as disinfection process.
Kim, Jin-Tae;Lee, Jong-Pil;Lee, Soo-Bin;Kim, Dong-Min
The Korean Journal of Nuclear Medicine Technology
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v.25
no.1
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pp.41-43
/
2021
Purpose In nuclear medicine blood tests, hemolysis samples are considered as inappropriate sample and are recommended not to be used for blood test. So, the lab are required to collect the blood again in the blood collection room However, The effect of hemolyzed samples on radioimmunoassay has not studied yet. This study was designed to evaluate effects of hemolysis on radioimmunoassay. Materials and Methods The kit manuals of 23 test items were reviewed to confirm whether hemolyzed samples were used. The subjects were 19 general applicants(male : 9, female : 13) and the samples were collected by each two SST tubes, one tube was obtained by centrifugation normally, and the other was obtained hemolyzed sample by centrifugation after external shock. It has been known that highly hemolyzed samples can affect the test results, so the test was performed using the severe hemolyzed sample. The test was performed for each test item using 23 normal serum and hemolysis serum, and SPSS19 program was used for statistical comparison of the test result. Results There was no significant difference between normal serum and hemolysis serum in 21 of 23 test items, but the results of insulin and C-peptide were significantly different(P<0.05). Conclusion It has been known that hemolysis in blood samples can affect the results of biochemical and hematological test, However, hemolysis effect is relatively low. Similarly, this study showed that hemolysis had not much effect on most of immunological radioimmunoassay except for some tests. Therefore, it is thought that the demand for re-collection due to hemolysis will be reduced in the laboratory, which will improve the work process of the laboratory.
Kim, Min-Ju;Kim, Byung-Hoon;Park, Sung-Soo;Park, Sung-Hee;Kim, Dong-Ho;Kim, Keun-Sung
Journal of Food Hygiene and Safety
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v.26
no.3
/
pp.198-202
/
2011
Rice has been the most important staple food in everyday meals of Korean people for thousands of years. Nowadays, it is getting increasingly used as flour ingredients in a variety of processed foods, so that it is consumed in more diversified ways. As a consequence, production volume of rice flour to manufacture rice cakes, noodles, breads, or confectioneries is recently getting increased in Korea. But there are not sufficient research outcomes to guarantee Korean consumers microbiological qualities of rice flour as well as rice. As a preliminary experiment, therefore, the microbiological profiles (aerobic mesophilic bacteria (AMB), spore-forming aerobic bacteria (SAB), lactic acid bacteria (LAB), yeasts and molds (YM), and Escherichia coli and coliforms) have been monitored for nine retailed white rice samples in this study. AMB counts ranged $10^2-10^6$ CFU/g for all the nine white rice samples. All the nine rice samples have SAB counts within a narrow range $(1.0{\times}10^2-2.5{\times}10^3$ CFU/g). LAB was detected in two white rice samples ($4.0{\times}10^2$ and $3.7{\times}10^3$ CFU/g), YM was detected in one white rice sample ($2.0{\times}10^2$ CFU/g) only. E. coli was not detected from all the nine samples. Coliforms were detected in one white rice sample ($4.1{\times}10$ CFU/g) only. All the rice samples were conclusively considered to have various microorganisms, though most of them are harmless and some, such as coliforms, may be harmful.
Purpose: Several studies report that detection of thyroglobulin (Tg) in fine-needle aspiration (FNA) biopsy washout fluid from lymph nodes identifies recurrences or metastases of differentiated papillary thyroid cancer (DPTC) in the neck with higher sensitivity and specificity than fine-needle aspiration cytology (FNAC). We evaluate the diagnostic efficacy and usefulness of Tg measurement in FNA washout fluid (FNA-Tg) and compare with FNAC. Materials and Methods: Forty-eight FNA samples of 37 patients who undergone ultrasonography to detect cervical lymph node metastasis of DPTC, were included for this study. Lymph node metastasis was confirmed by histopathologic examination or long-term imaging follow-up. Sensitivity, specificity and accuracy of FNA-Tg and FNAC were calculated. In 34 patients, we evaluated diagnostic accuracy of FNA-Tg according to the presence or absence of Tg antibody. Results: Sensitivity, specificity and accuracy of FNAC were 75.0%, 97.2% and 91.7%, respectively, and those of FNA-Tg were 100%, 88.9% and 91.7%, respectively. The presence of Tg antibody was not relevant to the diagnostic accuracy of FNA-Tg. Conclusion: FNA-Tg is a as accurate as FNAC with higher sensitivity. FNA-Tg and FNAC are complement techniques for diagnosing lymph node metastasis of DTPC.
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.
Although Mycobacterium tuberculosis complex strains remain responsible for the majority of diseases caused by mycobacterial infections worldwide, the increase in HIV (human immuno deficiency virus) infections has allowed for the emergence of other non-tuberculous mycobacteria as clinically significant pathogens. M. tuberculosis was detected by two-tube nested polymerase chain reaction (PCR) and non-tuberculous mycobacteria was detected by PCR-restriction fragment length polymorphism (RFLP) with Msp I. Result of niacin test is equal to result of two-tube nested PCR after culture for M. tuberculosis. In this study, acid fast bacilli stain (AFB. stain) >2+ case, Detection of Mycobacteria is similar to result before culture and after culture. AFB. stain <1+ case, result of mycobacteria is distinguished. Conclusionly, these results suggest that identification of mycobacteria must go side by side both culture and PCR for more fast and accuracy.
Park, Seung-Kyu;Kim, Seung-Chul;Kim, Deuk-Mi;Lee, Chang-Woon;Kim, Young;Cho, Sang-Nae
Tuberculosis and Respiratory Diseases
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v.53
no.6
/
pp.635-643
/
2002
Background : The Aim of this study was to compare the recovery of mycobacteria from sputum samples of pulmonary tuberculosis patients using the MB/BacT rapid culture system(Organon Teknika, USA) with that obtained using Lowenstein-Jensen solid medium. Methods : The two culture systems were compared using sputum samples of 99 pulmonary tuberculosis patients. Culture media were incubated at $35-37^{\circ}C$ for six weeks in the MB/BacT system and for 12 weeks in Lowenstein-Jensen solid medium. Solid media were examined macroscopically once a week, and the MB/BacT system positive vials were unloaded from the machine as soon as possible after positive signal from the connected computer was detected Confirmation of growth for mycobacteria was done by Ziehl-Neelson stained smears. Isolates were identified to differentiate Mycobacterium tuberculosis from mycobacterium other than tuberculosis(MOTT) by phenotypic and molecular methods. Results : Of the sputum samples of the 99 patients, 58 samples were smear positive and 41 in negative smear. Mycobacteria were recovered from 67(67.7%) samples by using both culture systems. The yield with MB/BacT was higher than that with Lowenstein-Jensen [67(67.7%) vs. 52(52.5%), p<0.001]. Moreover, 15(15.2%) samples were positive only in the MB/BacT, whereas none of samples was positive only in Lowenstein-Jensen. In smear-positive and smear-negative samples, the recovery rate with MB/BacT was also higher than that with Lowenstein-Jensen [sputum-positive; 56/58(96.6%) vs. 46/58(79.3%), p=0.005, sputum-negative; 6/41(14.6%) vs. 5/41(12.2%), p<0.001]. The mean times to detection of Mycobacteria were 13.3 and 27.2 days with MB/BacT and Lowenstein-Jensen respectively(p<0.001). Conclusion : This results indicate that the the MB/BacT is more efficient and faster than Lowenstein-Jensen for the recovery of mycobacteria.
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