Effectiveness of Real-Time Quantitative PCR Compare to Repeat PCR for the Diagnosis of Charcot-Marie-Tooth Type 1A and Hereditary Neuropathy with Liability to Pressure Palsies

The majority of cases of Charcot-Marie-Tooth type 1A (CMT1A) and of hereditary neuropathy with a liability to pressure palsies (HNPP) are the result of heterozygosity for the duplication or deletion of peripheral myelin protein 22 gene (PMP22) on 17p11.2. Southern blots, pulsed-field gel electrophoresis (PFGE), fluorescence in situ hybridization (FISH) and polymorphic marker analysis are currently used diagnostic methods. But they are time-consuming, labor-intensive and have some significant limitations. We describe a rapid real- time quantitative PCR method for determining gene copy number for the identification of DNA duplication or deletion occurring in CMT1A or HNPP and compare the results obtained with REP-PCR. Six patients with CMT1A and 14 patients with HNPP [confirmed by Repeat (REP) -PCR], and 16 patients with suspicious CMT1A and 13 patients with suspicious HNPP [negative REP-PCR], and 15 normal controls were studied. We performed REP-PCR, which amplified a 3.6 Kb region (including a 1.7Kb recombination hotspot), using specific CMT1A-REP and real-time quantitative PCR on the LightCycler system. Using a comparative threshold cycle (Ct) method and beta-globin as a reference gene, the gene copy number of the PMP22 gene was quantified. The PMP22 duplication ratio ranged from 1.35 to 1.74, and the PMP22 deletion ratio from 0.41 to 0.53. The PMP22 ratio in normal controls ranged from 0.81 to 1.12. All 6 patients with CMT1A and 14 patients with HNPP confirmed by REP-PCR were positive by real-time quantitative PCR. Among the 16 suspicious CMT1A and 13 suspicious HNPP with negative REP-PCR, 2 and 4 samples, respectively, were positive by real-time quantitative PCR. Real-time quantitative PCR is a more sensitive and more accurate method than REP-PCR for the detection of PMP22 duplications or deletions, and it is also faster and easier than currently available methods. Therefore, we believe that the real-time quantitative method is useful for diagnosing CMT1A and HNPP.

Utility of Tetanos Quick Stick(R) Test for Selective Tetanus Prophylaxis

PURPOSE: Tetanus is still one kind of major health problem in many countries, so tetanus prophylaxis is very important. However medical interview and wound description are not always enough to determine the tetanus prophylaxis. Thus, we assessed the utility of Tetanos Quick Stick(R) test for selective tetanus prophylaxis in the emergency department. METHODS: From September 17, 2003 to October 4, 2003, 180 patients were studied for 14 days. Tetanos Quick Stick (R) and ELISA (enzyme-linked immunosorbent assay) were performed with 180 samples from 180 patients. RESULTS: The Tetanos Quick Stick(R) had a sensitivity of 79.6%, a specificity of 99.2%, a positive predictive value of 97.7 %, a negative predictive value of 91.9%, and an accuracy of 93.3%. CONCILUSION: The results of our study revealed that the Tetanos Quick Stick(R) test is useful for selective tetanus prophylaxis in the emergency department.

Evaluation of Clinical Usefulness of PREG-Q(TM) as a Screening Test for Pregnancy / 임상검사와정도관리

BACKGROUND: Among the many methods estimating the quantity of beta-hCG for pregnancy testing in urine, immunochromatography is one of most widely used semi-quantitative detection method for its convenience to use and also for its rapid result reporting system. PREG-Q(TM) is a newly introduced semi-quantitative immunochromatography method for detecting b-hCG. Clinical usefulness of PREG-Q(TM) was evaluated as a screening test for early pregnancy detection. METHODS: Accuracy, detection limit, cross-reactivity with various glycoprotein hormones, interference study, and comparison study using total 100 urine samples from pregnant (50 samples) and non-pregnant women (50 samples) was evaluated. RESULTS: All the 50 urine samples of pregnant women showed positive results, and another 50 urine samples of non-pregnant women showed negative results with PREG-Q(TM). The lower detection limit of PREG-Q(TM) was 25 mIU/mL and the result was not affected by addition of glycoprotein hormones tested. Interfering substance causing false negative or false positive results enrolled didn't affect the test results in this study. CONCLUSIONS: We conclude PREG-Q(TM) is an excellent test kit for pregnancy test, and is valuable especially for detecting early pregnancy.

In Vitro Activities of Panipenem against Clinical Isolates of Aerobic and Anaerobic Bacteria

BACKGROUND: Panipenem is a carbapenem antimicrobial agent which has been shown to have broad-spectrum activities against various aerobic and anaerobic bacteria. In this study, in vitro activities of panipenem against recent clinical isolates of aerobic and anaerobic bacteria were determined. METHODS: Aerobic and anaerobic bacteria were isolated in 2001 and in 2000-2001, respectively, from a tertiary-care hospital patients. Antimicrobial susceptibility was tested by the NCCLS agar dilution method. RESULTS: MIC90s of panipenem were:similar to those of imipenem for aerobic gram-positive cocci and Enterobacteriaceae; slightly lower than those of meropenem for gram-positive cocci, but slightly higher for Enterobacteriaceae; slightly higher than imipenem for A. baumannii, but similar for anaerobic bacteria. CONCLUSION: MIC90s of panipenem were similar to those of imipenem for aerobic and anaerobic bacterial isolates, which frequently involve respiratory, urinary, intraabdominal and wound infections. When imipenem breakpoints are applied to interpret panipenem susceptibilities, panipenem can be considered useful for the treatment of various infections, including nosocomially acquired ones.

In Vitro Activities of Panipenem against Clinical Isolates of Aerobic and Anaerobic Bacteria

BACKGROUND: Panipenem is a carbapenem antimicrobial agent which has been shown to have broad-spectrum activities against various aerobic and anaerobic bacteria. In this study, in vitro activities of panipenem against recent clinical isolates of aerobic and anaerobic bacteria were determined. METHODS: Aerobic and anaerobic bacteria were isolated in 2001 and in 2000-2001, respectively, from a tertiary-care hospital patients. Antimicrobial susceptibility was tested by the NCCLS agar dilution method. RESULTS: MIC90s of panipenem were:similar to those of imipenem for aerobic gram-positive cocci and Enterobacteriaceae; slightly lower than those of meropenem for gram-positive cocci, but slightly higher for Enterobacteriaceae; slightly higher than imipenem for A. baumannii, but similar for anaerobic bacteria. CONCLUSION: MIC90s of panipenem were similar to those of imipenem for aerobic and anaerobic bacterial isolates, which frequently involve respiratory, urinary, intraabdominal and wound infections. When imipenem breakpoints are applied to interpret panipenem susceptibilities, panipenem can be considered useful for the treatment of various infections, including nosocomially acquired ones.

Two Cases of Cerebral Malaria Treated with Therapeutic Erythrocytapheresie / 대한수혈학회지

Cerebral malalria is a life-threatening complication of Plasmodium falciparum infection. RBC exchange transfusion (RCE) can reduce the burden of parasitemia in this situation. We have experienced two cases of cerebral malaria treated with automated RBC exchange as an adjunct to standard chemotherapy. Case 1: A 42-year-old male was referred to the emergency room with a history of 3 days of fever after having returned from Congo. Peripheral blood smear showed the P. falciparum parasitemia of 70-80%. Quinidine and doxycycline were administered but, mental state started to deteriorate. He underwent RCE on hospital day 2 to reduce the parasitemia to 10% after 8 hours. No parasite could be found on day 3 after the RCE. Case 2: A 62-year-old male was referred to the emergency room with a history of 3 days of fever after having returned from Cameroon. P. falciparum parasitemia was 10% on peripheral blood smear. Quinidine and doxycycline were immediately started but headache developed abruptly and he underwent RCE on hospital day 3. After 8 hours following the completion of RCE, parasitemia decreased to less than 1%. Automated RBC exchange transfusion can rapidly reduce the burden of parasitemia and achieve improvement of neurologic symptom and sign in patients with cerebral malaria.

BK Virus Detection by Polymerase Chain Reaction in Renal Transplant Recipients and Healthy Donors / 대한진단검사의학회지

BACKGROUND: BK virus is a polyomavirus associated with a range of clinical presentations from asymptomatic viruria with pyuria to ureteral ulceration with ureteral stenosis in renal transplant patients. BK viral Infection of renal allografts has been associated with diminished graft function in some individuals. We tried to detect BK virus in urine and plasma from Korean renal transplant recipients, renal transplant candidates, and healthy donors. METHODS: To detect BK virus in urine and plasma, we used PCR-RFLP (polymerase chain reaction and restriction fragments length polymorphism) with BamHI. The study was performed from 118 renal transplant recipients, 18 renal transplant candidates, and 25 healthy donors. RESULTS: BK virus DNAs were detected in 21.2% of urine and 0.9% of plasma from renal transplant recipients. BK virus DNA was detected in neither urine nor plasma from healthy donors and renal transplants candidates. Among a total of eight patients who were clinically suspected of having BK nephropathy, three were PCR positive for BK virus and two were decoy-cell cytology positive. Six patients were diagnosed as BK nephropathy by tissue pathology. Among them, BK virus was detected by PCR in urine from five patients, and decoy cells were shed from five patients, respectively. CONCLUSIONS: BK virus detection by polymerase chain reaction in urine may be a non-invasive and sensitive tool for diagnosing and monitoring BK nephropathy.

An Experience of Cord Blood Transfusion in Cold Hemagglutinin Disease Associated with Mycoplasma pneumoniae Infection / 대한진단검사의학회지

We report a case of cold hemagglutinin disease associated with Mycoplasma pneumoniae infection treated with cord blood transfusion. Cold hemagglutinin disease is a hemolytic anemia most commonly associated with cold-reactive autoantibody with anti-I specificity. On the basis of the fact that the level of I antigen on cord red blood cells is extremely low, a six year old male patient was transfused with 60 mL of ABO blood type-matched, cord blood. No complication from the transfusion was observed. Due to the deficiency in cord blood supply, filtered irradiated RBC 100 mL was transfused three times thereafter. The hemoglobin level began to increase from the fifth hospital day. The patient was discharged without additional transfusion on the eleventh hospital day. No remarkable complications were noted at the time of discharge.

Red Cell Depletion from the Bone Marrow Aspirates for the ABO Incompatible Transplantation by Apheresis Separations / 대한혈액학회지

BACKGROUND: Allogeneic bone marrow transplantation across ABO incompatibility barriers may result in immune mediated hemolysis. Hemolysis may be avoided by RBC depletion from the graft. In vitro graft manipulations carry the risk of hematopoietic stem cell loss, a factor that may be most important in graft failure. We report 16 major ABO blood group incompatible allogeneic bone marrow transplants using erythrocyte depletion of marrow prior to infusion. METHODS: From March 1997 to July 2001 in Yonsei University College of Medicine, 16 patients underwent ABO blood group incompatible allogeneic BMT: 5 for acute myelocytic leukemia, 5 for severe aplastic anemia, 3 for acute lymphocytic leukemia, 2 for chronic myelocytic leukemia, and 1 for myelodysplastic syndrome. RBC depletions were done with automatic cell separator, COBE Spectra (COBEBCT Inc., Lakewood, USA). RBC removal rates and mononuclear cell recovery rates were calculated. And the evidence of successful engraftment and intravascular hemolysis were also evaluated. RESULTS: The RBC removal rate was 99.1+/-0.0% and a mean of 1% of the original red cell volume was contained in the final infusate. The mononuclear cell recovery rate was 70.0+/-16.3% from the original MNCs. Fourteen patients tolerated the infusion of the marrow concentrates without any adverse effects. Two patients experienced hemoglobinuria, but disappeared within 2 days by continued observation. After transplantation, absolute neutrophil counts exceeded 500/nL by 10.8+/-1.9 days, platelet counts exceeded 50,000/nL by 30.5+/-8.5 days, and reticulocytosis sustained at >1% was by 25.8+/-13.9 days. CONCLUSION: RBC depletion from ABO major mismatched bone marrow aspirates by the automatic cell separator is a safe and effective technique.

A Survey of Blood Component Use in Relation to International Statistical Classification of Diseases (ICD)-10 / 대한임상병리학회지

BACKGROUND: There are few recent studies on transfusion practice and blood use with regard to diagnoses of Korean recipients. We conducted a survey of blood component use in Yonsei University Medical Center in Seoul, Korea. METHODS: A survey was carried out on blood component transfusion from January 2000 to June 2000. Transfused components were listed by broad diagnostic categories formed from the principal diagnoses of the recipients according to the ICD-10. RESULTS: Of all investigated components (67,433 units), 18,623 units of RBCs (27.6%), 10,203 units of FFP (15.1%), 38,547 units of platelets (57.2%), and 60 units of whole blood (0.1%) were transfused. The transfusion rate in relation to sex was 1.7:1 and the majority of all blood units were transfused to patients aged <65 yrs (85.9%). The rate of blood component transfusion was the highest in the department of internal medicine (47,451 units, 70.4%). Of 18,623 RBCs units and 10,203 FFP units, 70.8% and 74.5% were transfused to patients in four diagnostic categories; neoplasms, digestive system diseases, circulatory system diseases, and leukemia or lymphoma. Of 38,547 platelets units, 87.8% were used in patients with neoplasms, leukemia or lymphoma, digestive system diseases and diseases of the blood and blood-forming organs. CONCLUSIONS: This survey provides information on the blood component usage in tertiary-care teaching hospital in Seoul, Korea. It demonstrates the concentration of today's blood utilization among a few diagnostic categories. This information is relevant for quality management of transfusion practice, cost analyses and for planning local and regional blood donation programs.

The Incidence of Unexpected Antibodies in Transfusion Candidates / 대한수혈학회지

BACKGROUND: Some investigators have done several studies on the frequency and distribution of unexpected antibodies in Korea. But more studies are necessary to estimate the precise frequency of unexpected antibodies. METHOD: In order to determine the frequency of unexpected antibodies, we analyzed the results from 23,735 antibody-screening tests performed for transfusion candidates at Yonsei University Severance Hospital during a recent two-year period (March 1998~ Feb 2000). Screening and identification of unexpected antibodies were carried out using a gel agglutination technique with DiaMed-ID system (DiaMed, Murten, Switzerland). RESLUT: Out of all 23,735 serum samples, positive results were obtained from 109 sera (0.46%). Clinically significant antibodies were found from 0.29% of all the population. Antibodies that were detected most frequently were anti-E+c (33 sera), followed by anti-E (19 sera) and anti-Lea (12 sera). CONCLUSION: We found that Rh antibodies, clinically significant unexpected antibodies, are detected frequently in Korea, too. So we consider that unexpected antibody screening test must be included in pre-transfusion test in Korea and this information is reassuring in assessing the risk associated with blood transfusion in many hospitals of Korea.