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Gender Verification Test Based on PCR Determination of Y Chromosomal DNA: Experience in Genetic Sex Typing during the '99 Kangwon Asian Winter Games.

Jeong Uk KIM ; Hwan HERR ; Hyun Kyu CHANG ; Hae Kyoung AHN ; Kil Joo OH ; Hong Ki KIM

Korean Journal of Clinical Pathology.2000;20(2):215-246.

BACKGROUND: The aim of gender verification test is to maintain impartiality among female competitors by excluding males in women's sports competitions. Some microscopic methods such as X-chromatin test and Y-chromatin test had been used for this purpose. Because of their known shortcomings, the methods were replaced with the polymerase chain reaction(PCR)-based test. In this report we describe the assay used in the gender verification during the '99 Kangwon Asian Winter Games. METHODS: Buccal smear samples of 126 female competitors were obtained. These samples underwent digestion with proteinase K, and were followed by boiling treatment with Chelex resin. PCR was performed to detect the sex determining region of Y chromosome(SRY) in order to confirm the femininity, and beta globin region was coamplified for confirming that the DNA was extracted from buccal cells. An X-Y homologous region encoded amelogenin was also amplified so that the femininity could be reconfirmed. RESULTS: No SRY and Y-amelogenin like sequences were amplified in any of samples of 126 female competitors analysed. CONCLUSIONS: Established gender verification method based on PCR amplification of Y chromosomal DNA seems to be superior to others. Sampling is simple. The procedure of extracting DNA is simple, rapid, and does not require multiple tube transfers. False positivity and/or false negativity appear to be less. It appear that this method is useful and reliable for gender verification in international sports events.
Amelogenin ; Asian Continental Ancestry Group* ; beta-Globins ; Digestion ; DNA* ; Endopeptidase K ; Female ; Femininity ; Gangwon-do* ; Humans ; Male ; Polymerase Chain Reaction* ; Sports

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Positive Rate of HLA Class I Antibodies in Multiparous Korean Women.

Eun Young SONG ; Sun Mee KIM ; Byoung Cheol KIM ; Kyou Sup HAN ; Myoung Hee PARK

Korean Journal of Clinical Pathology.2000;20(2):210-214.

BACKGROUND: Anti-HLA antibodies are most frequently induced by transfusion or pregnancy, and these anibodies can be used as antisera for HLA typing. However these antibodies may elicit adverse reactions such as transfusion reaction or rejection of transplanted organs. In this study, frequency and specificities of antibodies against HLA class I antigens were determined in multiparous Korean women. METHODS: Sera from 671 multiparous women were tested for anti-HLA antibody screening by standard microlymphocytotoxicity test using 49~50 lymphocyte panels. PRA(panel reactive antibody) values were calculated as percentage of postive panels among total lymphocyte panels tested. HLA antibody specificities and reaction strengths were determined by analysis of serologic reaction patterns. RESULTS: A total of 671 sera were tested and 124 sera(18.5%) were positive for HLA antibodies. Among HLA antibody-positive sera(n=124), 117(94.4%) showed PRA values of < OR = 50% and only 7(5.6%) showed PRA values of >50%. Specificities of HLA antibodies were identified in 51 sera(41.1%) and 18 sera(14.5%) contained reagent quality antibodies(r> or =0.8, SI> or =70%), corresponding to 2.7% of total multiparous women. Among these, 4 sera had monospecific HLA antibodies and 14 sera had HLA antibodies against two or more antigens: 4 sera containing HLA antibodies against 7 CREG(cross reactive group), 5 sera containing antibodies against 5 CREG. CONCLUSION: Through the analysis of frequency and specificity of HLA antibodies in 671 multiparous women, it is concluded that HLA antisera can be obtained from multiparous women as effectively as from pregnant women. The frequency of high level of sensitization(PRA>50%), which can elicit problems in relation to transfusion or organ transplantation, is very low(1.0%).
Antibodies* ; Antibody Specificity ; Blood Group Incompatibility ; Female ; Histocompatibility Antigens Class I ; Histocompatibility Testing ; Humans ; Immune Sera ; Lymphocytes ; Mass Screening ; Organ Transplantation ; Parity ; Pregnancy ; Pregnant Women ; Sensitivity and Specificity ; Transplants

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Frequency and significance of concurrent presence of HBsAg and anti-HBs among Korean HBsAg-positive patients.

Young Joo CHA ; Seok Lae CHAE

Korean Journal of Clinical Pathology.2000;20(2):204-209.

BACKGROUND: Although the development of antibodies(anti-HBs) against hepatitis B surface antigen(HBsAg) generally leads to the clearance of the infecting hepatitis B virus(HBV), anti-HBs reactivity has been reported in patients with chronic hepatitis. Some reports suggested that concurrent presence of HBsAg and anti-HBs was a common serologic pattern in HBV carriers, however, the others insisted concurrence was associated with evidence of viral replication and features of active inflammation. This study was to investigate the frequency and significance of concurrent presence of HBsAg and anti-HBs among Korean HBsAg-positive patients. METHODS: HBV serological markers were analyzed by Abbott Axym. Anti-HBs titer was assessed quantitatively using 6 standard calibrators(0, 10, 50, 100, 500 & 1,000 mIU/mL), and reported as positive when more than 10 mIU/mL. RESULTS: Of 959 consecutive HBsAg-positive patients who had simultaneous anti-HBs determinations, HBsAg and anti-HBs were found concurrently in 31(3.2%). Anti-HBs was quantitated 10-100 mIU/mL in 25(80.6%) and more than 100 mIU/mL in 6(19.4%) among 31 HBsAg-positive patients with concurrent presence of anti-HBs. HBeAg positive rate of concurrent HBsAg/anti-HBs-positive patients(15/22 or 68.2%) was higher than that of HBsAg-positive patients without anti-HBs(227/562 or 40.4%). Of 31 HBsAg-positive patients with concurrent anti-HBs, 15 were patients with chronic hepatits, 5 with primary hepatocellular carcinoma and 2 with liver cirrhosis. CONCLUSIONS: Concurrent HBsAg/anti-HBs-positive patients were found in about 3% of Korean HBsAg-positive patients and their anti-HBs titers were less than 100 mIU/mL in 80% of concurrent HBsAg/anti-HBs-positive patients. In patients with chronic hepatits B who possessed HBsAg and anti-HBs above 100 mIU/mL, infections of HBV variants with mutations in the envelope gene might be suspected.
Carcinoma, Hepatocellular ; Hepatitis B ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens* ; Hepatitis, Chronic ; Humans ; Inflammation ; Liver Cirrhosis

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Comparison of HLA-B27 typing methods -PCR-SSP, microlymphocytotoxicity, and flow cytometry.

Jun Soo BAE ; Young Ree KIM ; Hyun Il CHOI ; Yun Jung CHO

Korean Journal of Clinical Pathology.2000;20(2):198-203.

BACKGROUND: HLA-B27 typing has long been performed by the microlymphocytotoxicity method(MCT) but the flow cytometry method(FCM) was introduced several years ago. False positive results due to the HLA-B7 cross reactive groups(CREG) were the main drawback of the serologic method. The authors performed polymerase chain reaction-sequence specific primer(PCR-SSP) test for HLA-B27 to compare the results with serologic methods. METHODS: PCR-SSP test for HLA-B27 was performed on four hundred forty one samples. Three hundred twenty eight samples were tested by MCT and one hundred thirteen samples by FCM. PCR-SSP for HLA-B27 subtyping or Amplification Refractory Mutation System-PCR(ARMS-PCR) for HLA-B typing was performed on twenty four discrepant samples. RESULTS: The concordance rate between MCT and PCR-SSP was 92.9%(305/328) and the concordance rate between FCM and PCR-SSP was 99.1%(112/113). Twenty four(5.4%) out of four hundred forty one samples showed discrepancy between serologic methods and PCR-SSP method. Fourteen out of one hundred MCT positive samples and only one out of forty FCM positive samples showed negative by PCR-SSP. Nine samples showed PCR-SSP positive and MCT negative. CONCLUSIONS: The false positive rate of MCT was quite high and there were some false positive and negative results by PCR-SSP, too. From the above findings, we suggest that FCM is the most accurate method for HLA-B27 typing in those laboratory equipped with flow cytometry.
Flow Cytometry* ; HLA-B Antigens ; HLA-B27 Antigen* ; HLA-B7 Antigen

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A Case of Septicemia without Skin Lesion by Serovar O1 Type of Vibrio vulnificus Infection.

Jeong Ill SUH ; Kyung Im HA ; Yunsop CHONG

Korean Journal of Clinical Pathology.2000;20(2):194-197.

Vibrio vulnificus infections occur between May and October, and most often involve men over age 40. V. vulnificus has been linked to two distinct syndromes: primary sepsis and primary wound infections. Primary sepsis occurs most commonly in patients with antecedent liver disease such as cirrhosis. Mortality approaches about 50 percent, with most deaths due to uncontrolled sepsis. We experienced a case of septicemia without dermatologic lesion by serovar type O1 V. vulnificus, and reported this case with a review of literature.
Fibrosis ; Humans ; Liver Diseases ; Male ; Mortality ; Sepsis* ; Skin* ; Vibrio vulnificus* ; Vibrio* ; Wound Infection

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Evaluation of Mycobacterial Recovery by Specimen Preparation and Inoculating Media.

Joseph JEONG ; Chulhun L CHANG

Korean Journal of Clinical Pathology.2000;20(2):188-193.

BACKGROUND: During the specimen preparation, neutralization after alkali processing is not necessary when inoculate into agar or broth media, but into 3% Ogawa media, the mostly used egg-based media in Korea. To simplify the specimen processing, the recovery of mycobacteria from 3% Ogawa media inoculating differently processed sputa, was evaluated. Futhermore, the recovery from the three different media was evaluated, too. METHODS: 209 sputa were included. Each specimen was divided into 3 bottles.(A) One was only decontaminated by 4% NaOH before inoculation.(B) Another was decontaminated, and concentrated without neutralization. These two specimens were inoculated into 3% Ogawa media.(C) The other was decontaminated, neutralized and concentrated, and innoculated into 3% Ogawa, Middlebrook 7H9 selective broth and 7H10 selective agar media. The isolates were identified by AccuProbe method. RESULTS: M. tuberculosis were isolated from 43(20.5%) specimens. The recovery number and time from Ogawa media, inoculating specimens processed by A, B, and C method, were 29(13.9%), 35(16.7%), and 36(17.2%), and 23, 23, and 19 days, respectively. The recovery number and time from Ogawa, 7H10 agar and 7H9 broth media, inoculating specimens processed by C method, were 36(17.2%), 42(20.1%), and 41(19.6%), and 26, 24, and 22 days, respectively. CONCLUSIONS: Neutralization of alkali-decontaminated specimens could be used to inoculate into 3% Ogawa media. Inoculation into homemade 7H10 agar and 7H9 broth media in addition to 3% Ogawa media improved the recovery rate and time of mycobacteria.
Agar ; Alkalies ; Korea ; Mycobacterium tuberculosis ; Sputum ; Tuberculosis

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A Case of Transient Myeloproliferative Disorder in Robertsonian Translocation Type of Down's Symdrome.

Gueong Seon KIM ; Chun Hee LEE ; Young Sook KIM ; Oh Kyung LEE

Korean Journal of Clinical Pathology.1997;17(4):530-538.

A case of Down's syndrome in which transient myeloproliferative disorder developed is described. In hematologic findings of peripheral blood, high blast cell count on 1st day of birth had been found and after serial follow-up for several weeks, decrease in WBC counts from 22.6x10(9)/L to 7.5x10(9)/L and blast cell counts from 31% to 2% occurred. The karyotype of his patient was 45,XY, der (13;14) (q10;q10), der(14;21) (q10;q10), +21. Karyotyping of his father revealed 45,XY, der(13;14) (q10;q10). Without specific chemotherapy, hematologic and clinical recovery was occurred within several weeks. We deport a case of transient myeloproliferative disorder in Robertsonian translocatlon type of Down's syndrome accompanying another Robertsonian translocation, der (13;14) (q10;q10), inherited from his father.
Cell Count ; Down Syndrome ; Drug Therapy ; Fathers ; Follow-Up Studies ; Humans ; Karyotype ; Karyotyping ; Myeloproliferative Disorders* ; Parturition

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Acute Lymphoblastic Leukemia with Philadelphia Chromosome and Monosomy 7.

Gui Jeon CHOI ; Hyo Jin CHUN ; Dong Seok JEON ; Jae Ryong KIM ; Myung Soo HYUN

Korean Journal of Clinical Pathology.1997;17(4):519-529.

BACKGROUND: The combination of Philadelphia chromosome (Ph) and monosomy 7(-7) was rarely observed in acute lymphoblastic leukemia (ALL). With the results from immunophenotyplc and molecular analysis, Philadelphia chromosome positive ALL with monosomy 7[Ph(+)/-7] has been considered that it may be derived from neoplastic transformation at the pluripotent stem cell level. We compared the clini-cal, laboratory, and hematological findings between 5 cases of Ph(+)/-7 and 5 cases of Ph(+) without monosomy 7 [Ph (+) /N7]. METHODS: During the period from January, 1995 to December, 1996, total 72 cases of ALL were confirmed among 259 cases of hematologic malignancy with bone marrow cytogenetic analysis. Among 72 ALL cases, 5 cases of Ph(+)/-7(monosomy 7 or 7q abnormalities) were compared with Ph only or Ph without monosomy 7(ph(+)/N7] on the hematological, immunophenotypic, other laboratory, clinical findings and event ree survival (EFS) The karyotyping of the bone marrow specimens was analysed byshort-term unsynchronized culture methods such as overnight colcemid treatment and 24 hours incubation following ethidium bromide treatment. RESULTS: The mean age of Ph(+)/-7 was 30.6+/-12.8 years, and it was significantly different from that of Ph(+)/N7 (p=0.009), Four cases of Ph(+)/-7 were classified as ALL L2 subtype, and 2 cases revealed CNS involvements. Immunophenotyping was positive in CD10, CDl9, CD2O, CD22 and HLA-DR. But one case revealed e-B-lymphoid lineage with positivity in CD34, CDl3, and CD33. The response to chemotherapy and EFS was very poor in Ph(+)/-7 group, and the mean EFS was 3.2+/-1.9 months(p=0.014). All of cases showed induction on failure in chemotherapy, relapsed with bone marrow, CNS and extramedullary involvements, and expired due to sepsis. CONCLUSIONS: Ph(+)/-7 ALL had very Poor clinical course with being resistant to chemotherapy and unfavorable prognosis, revealed L2 subtype by FAB classification, and was slightly older in ages compared with Ph(+)/N7 ALL.
Bone Marrow ; Classification ; Cytogenetic Analysis ; Demecolcine ; Drug Therapy ; Ethidium ; Hematologic Neoplasms ; HLA-DR Antigens ; Hydrogen-Ion Concentration ; Immunophenotyping ; Karyotyping ; Monosomy* ; Philadelphia Chromosome* ; Pluripotent Stem Cells ; Precursor Cell Lymphoblastic Leukemia-Lymphoma* ; Prognosis ; Sepsis

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Chromosome Breakage Test for the Diagnosis of Fanconi's Anemia.

Dong Wook RYANG ; Deok CHO ; Won Pyo HONG ; Hoon KOOK ; Tai Ju HWANG

Korean Journal of Clinical Pathology.1998;18(1):101-106.

BACKGROUND: Fanconi's anemia (FA) is an autosomal recessive disease characterized by aplastic anemia, pre-malignancy, congenital malformations and chromosome breakage syndromes. As up to 30% of patients have no detectable congenital anomalies, the modern diagnosis of FA rests on chromosomal breakage of patient's cells induced by chemical clastogens such as diepoxybutane (DEB) or mitomycin-C (MMC). METHODS: We have done chromosome breakage test to differentiate FA from 11 aplastic anemia, three Diamond-Blackfan syndrome, three myelodysplastic syndrome, one acute leukemia with congenital anomaly and three siblings of FA. The peripheral blood lymphocytes from each individual were co-cultured in phytohemagglutinin-containing medium by the three methods, i.e., DEB treated, MMC treated and un-treated. RESULTS: Five cases were found to have increased chromosomal breakages to DEB and MMC, confirming diagnosis of FA. Other 21 cases showed no increased chromosomal breakages. No overlap was found between FA group and others (P<0.01). In one FA, there was no increased spontaneous breakage, but increased breakage to DEB and MMC. Of five FA, one case showed no congenital anomalies. CONCLUSIONS: Chromosme breakage test was shown to be simple, reliable and useful in ascertaining the diagnosis of FA.
Anemia, Aplastic ; Chromosome Breakage* ; Diagnosis* ; Fanconi Anemia* ; Humans ; Leukemia ; Lymphocytes ; Mitomycin ; Mutagens ; Myelodysplastic Syndromes ; Siblings

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The Re-evaluation of the Serologic ELISA tests for the Diagnosis of Helicobacter pylori Infection.

Sinkyung KIM ; Think You KIM

Korean Journal of Clinical Pathology.1998;18(1):96-100.

BACKGROUND: Helicobacter pylori (H. pylori), a Gram negative spiral bacilli, is known to be the cause of chronic gastritis and peptic ulcer disease and is strongly associated with gastric cancer. Therefore, the rapid detection of H. pylori infection is necessary for prevention and treatment. Of the diagnostic tests currently used, the serologic tests, which makes use of the immune response, do not need a biopsy specimen. This method is relatively accurate, rapid, simple and inexpensive. We re-evaluated the clinical usefulness of the isotypes of H. pylori (IgG, IgA, IgM) antibodies for the detection of H. pylori infection. MATERIALS AND METHODS: Serum samples were obtained from 1,851 patients confirmed to have gastric or duodenal disease by gastric endoscopy from June, 1993 to December, 1994 at Hanyang University Hospital. The phenol-red urease test was done during endoscopy and the H-E stain on the gastric biopsies. Serologic tests (GAP Test IgG, IgA, IgM kits) were performed with patient sera. RESULTS: The sensitivities of the GAP EIA were 80% for IgG, 27% for IgA, and 85% for IgM. The specificities were 33%, 79%, and 14%, respectively. The detection rates of H. pylori were highest for the phenol-red urease test (88%), followed by IgM by ELISA (86%), IgG (72%), H-E stain (43%), and IgA (21%). The serum levels of IgG and IgA antibodies were higher in those with H. pylori infection than in those without, but there was no difference in IgM levels. And, no difference of serologic antibody levels according to disease state. Where follow-up was possible, the majority of IgG levels decreased, but IgA or IgM levels are not changed. CONCLUSION: A positive serologic test is incapable of discriminating between actual infection and normal bacterial colonization, or between recent and past infections. Therefore a serologic test seems unsatisfactory for confirming a diagnosis of H. pylori infection, but because the serial IgG levels of treated patients decreased significantly, we believe that this test may be used as an indirect means of assessing the response to therapy.
Antibodies ; Biopsy ; Colon ; Diagnosis* ; Diagnostic Tests, Routine ; Duodenal Diseases ; Endoscopy ; Enzyme-Linked Immunosorbent Assay* ; Follow-Up Studies ; Gastritis ; Helicobacter pylori* ; Helicobacter* ; Humans ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Peptic Ulcer ; Serologic Tests ; Stomach Neoplasms ; Urease

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