We mainly depend on the eyes to discriminate colors and the evaluation thus becomes subjective. The development of the spectrophotometer, however, allows us to analyze colors objectively. In this study, we tried to evaluate objectively the stainability of the hematoxylin–eosin (HE) stain using this instrument. Specimens stained with the HE stain were collected from 18 laboratories, which were members of the Study Group for Pathology and Cytology Laboratory, Aichi Association of Laboratory Technologists, and their stain densities were analyzed using a spectrophotometer (Konica-Minolta CM5, Japan). In addition, 18 hematoxylin solutions as well as 18 eosin solutions were divided into 3 categories according to their color densities, namely, dark, intermediate, and light. Then, we paired each one of the 18 hematoxylin solutions with 3 different densities of eosin solutions, and in the same way, each one of the 18 eosin solutions with 3 different densities of hematoxylin solutions. The results showed that the main factor contributing to the effect on the stainability of the HE stain was not the density of the hematoxylin solution but rather that of the eosin solution. This finding suggests that the control of the density of the eosin solution could be the main factor for determining the final result of the HE stain. We consider that adjusting the density of the eosin solution might be the most important key element for the standardization of the HE stain.

View full abstract

We have examined the differences in staining in HER2 tests (HER2-IHC and HER2-FISH) with different fixatives using a transplant xenograft model mouse producing c-erbB-2 (4-1ST tumor line). The tissue fixatives use 10% and 20% non-buffered formalin, 10% and 20% neutral buffered formalin solution. The HER2-IHC test was performed on a tumor tissue fixed for 4, 48, 72 and 168 h. The HER2-IHC kits used six commercial products and the HER2-FISH kits used one product. The results of the HER2-IHC tests showed strong positive (score of 3+) findings with fixation times of 4–168 h. However, for the sample that is fixed for 168 h in 20% non-buffered formalin, the staining of the cell membrane is attenuated by one product. The HER2-FISH test for the confirmation of the overexpression of the HER2 gene and CEP17 was confirmed in all fixative materials. The 4-1ST xenograft tumor is not affected by the concentration and type of fixative, which proved that the tumor antigen protein or the DNA is maintained. 4-1ST xenograft model mice are useful as external quality assessment samples required for quality control in HER2 testing.

View full abstract

Moraxella catarrhalis is present in the upper respiratory tract often together with Streptococcus pneumoniae and Haemophilus influenzae and it causes sinusitis, tympanitis and other respiratory ailments. The national vaccination program in Japan against S. pneumoniae and H. influenzae was revised in 2010 so that children less than 5 years of age are eligible for vaccination. It is, therefore, potentially conceivable that the isolation frequencies of these pathogens may have varied thereafter. We conducted a survey of the occurrence of M. catarrhalis for six consecutive years from 2009 to 2014 at Kitasato University Hospital. The isolation frequency of M. catarrhalis from 2009 to 2014 increased consistently with age. This trend was markedly significant in children up to 5 years of age, suggesting a possibility that the vaccination program affected the populations of these pathogens. Among M. catarrhalis isolates, the MIC₉₀ of levofloxacin-resistant strains increased 2-fold in children less than 5 years of age and 4-fold in adults more than 65 years of age in 2013. We interpreted the data to mean that the levofloxacin-resistant strains had been transmitted from elderly carriers to these children, because new quinolone antibiotics are not prescribed to children less than 5 years of age in general.

View full abstract

Severe fever with thrombocytopenia syndrome (SFTS) was first defined as a new tick-borne infectious disease in China in 2009. In this study, the sensitivities of detection of SFTS virus with two primer sets in the polymerase chain reaction (PCR) were examined. Blood samples were obtained from 52 cases with suspected SFTS from March 2013 to March 2015 in Ehime prefecture, Japan. Of these samples, 17 were positive in PCR, and 16 of the 17 positive samples were collected by the sixth day after onset of the disease, suggesting that blood samples should be collected as early as possible after onset. The detection rate was 77.8% (14 of 18 cases) with primer set 1 and 94.4% (17 of 18 cases) with primer set 2. In 1 of the 3 cases that were negative with primer set 1 but positive with primer set 2, a single-nucleotide mismatch (A→G) for the NP-1F primer of set 1 was present in the position sixth from the 3' end. Therefore, the detection sensitivity of PCR for SFTS virus may be affected by PCR conditions, such as primer-template mismatches and template amounts. Such factors may have caused the lower detection rate with primer set 1 than with primer set 2. Phylogenetic analysis of the NP region (420 bp) revealed that the Ehime strains of SFTS virus belonged to a cluster with previously reported Japanese strains, which are distinct from Chinese strains.

View full abstract

A medicinal toxicant is sometimes the cause of consciousness disorder and shock symptoms in a patient at the site of emergency medical care. A rapid drug screening kit is used as a measure for determining the cause. INSTANT-VIEW M-I (hereinafter referred to as “INSTANT-VIEW”), which is distributed by FUJIREBIO INC. in Japan, can detect 6 types of drug or drug metabolite at the same time, namely, stimulant (METH), cannabis (THC), cocaine (COC), benzodiazepine (BZD), barbiturate (BAR) and tricyclic antidepressants (TCA). The assay kit with a one-step method is easy to use and the reaction time is short at only 7 minutes. However, the operator should pay attention because the interpretation becomes positive with linear disappearance, and the interpretation with a linear disappearance is opposite to that obtained using a general ‎immunochromatographic assay kit. We confirmed that some drug samples have a density even more than tenfold that described in the package insert, and these could not be detected owing to the measurement detection limit using imitation urine samples, in which a standard solution is added to a healthy control urine sample. Tetracyclic antidepressants and antipsychotics as TCAs indicate a false positive result following a cross-reactive examination. It is important to understand that there is a possibility of obtaining false positive and false negative results during the interpretation of the results of immunochromatographic assay kits including INSTANT-VIEW.

View full abstract

Methotrexate (MTX), regarded as an anti-metabolite and anti-folate drug, is used in the treatment of cancer and autoimmune diseases such as rheumatoid arthritis. Since high-dose MTX is especially effective for the treatment of acute leukemia and malignant lymphoma, measuring MTX is indispensable to avoid adverse reactions and to complete efficient leucovorin rescue therapy. The purpose of this study was to evaluate the analytical performances of Nanopia eTDM Methotrexate and ARCHITECT Methotrexate in comparison with EMIT Methotrexate in terms of reproducibility, dilution linearity, detection limits, interference, DAMPA cross-reactivity, and correlation. The accuracies of these 3 assays were also evaluated with prepared samples, the concentrations of which were determined by gravimetric method. The ARCHITECT Methotrexate assay showed the best analytical performance among the assays evaluated. Because of its accuracy at low concentration and rapid measurement, the ARCHITECT Methotrexate assay is a useful method for therapeutic drug monitoring.

View full abstract

Liver-type fatty acid binding protein (L-FABP) is a 14 kDa protein expressed in the cytoplasm of human renal proximal tubule cells. Renal L-FABP expression is up-regulated and the urinary excretion level of renal L-FABP is increased by various stressors, such as hyperglycemia, urinary protein, and tubular ischemia. Therefore, the measurement of urinary L-FABP is useful for the early diagnosis of kidney disease with renal tubular disorders. In our study, we evaluated the basic performance of “Nordia^®L-FABP” for the measurement of L-FABP in human urine by latex turbidimetric immunoassay. The result indicates that the variation coefficients for within-run and day-to-day run reproducibilities were 1.3 to 2.2% and 1.2 to 3.9%, respectively. Furthermore, the linearity was also tested by serial sample dilution and was observed up to a dilution of 210 ng/mL, and the detection limit was 1.2 ng/mL. Compared with the conventional reagent, it has sufficient basic performances, and the correlation test was good (y = 1.12x + 0.53, r = 0.976). There was no significant interference caused by the addition of the following interfering substances: conjugated bilirubin, hemoglobin, and chyle. On the other hand, ascorbic acid may affect the measured levels of L-FABP depending on the concentration of coexisting substances. These results indicate that the new method, applicable to automated clinical chemistry analyzers, is useful for rapid measurement of L-FABP in human urine, and this assay kit may be beneficial in routine tests because it is useful for the early diagnosis of kidney disease.

View full abstract

A seventy-one-year-old male was transferred to the emergency room of our hospital with the diagnosis of suspected meningitis by a previous physician owing manifestations of sudden-onset high-grade fever, unconsciousness, and nuchal rigidity. On spinal tap examination, microscopy observation showed numerous bacteria and no pleocytosis, and a chemical examination revealed a low glucose level and high levels of protein, lactate dehydrogenase (LD), and creatine kinase (CK). LD isozyme analysis detected LD1, LD2, and LD3, which come from brain cells, but not LD4 or LD5. CK isozyme analysis showed an increase in the level of CK-BB, indicating brain cell damage or brain tissue injury. Unfortunately, he died on the same day. Streptococcus agalactiae cells were cultured from his blood and spinal fluid samples. Thus, he was suspected as having had bacterial meningitis. This patient was considered immunocompromised because of other underlying diseases or physiology such as being elderly and having diabetes mellitus and chronic renal failure. Because of his poor immune response, the patient showed no pleocytosis, and severe bacterial infection and brain cell damage developed rapidly.

View full abstract

Intervention by clinical laboratory staff members in the evaluation of non-negligible false values in clinical laboratory tests can lead to appropriate management of patients. Serum/plasma potassium levels can be influenced by pre-analytical processes, and falsely elevated potassium levels can mislead physicians to prescribe unnecessary treatments. A 72-year-old woman with autoimmune hepatitis was hospitalized. In the examinations during hospitalization, her potassium levels varied (3.9–6.2 mmol/L) but her condition was consistent. Her laboratory findings showed no evidence of hemolysis, and there was no marked increase in blood cell counts. To determine the condition of blood collection, we visited her room. Visiting her bedside revealed that her handgrip was excessive before blood collection. Her potassium level after sufficiently opening her hand was 4.0 mmol/L, whereas that after excessive handgrip was 4.9 mmol/L. Thus, her potassium levels could be affected by the excessive handgrip. To share information on blood collection with doctors and nurses, these findings were written in the medical record and she was educated to tell blood-collection operators about the potential elevation of potassium levels when her blood is drawn. After the information sharing and patient education, no elevated potassium levels were observed. The quality control of pre-examination processes requires not only measures by medical staff members but also self-awareness on the part of the patient. Active cooperation and information sharing by clinical laboratory staff members can contribute to achieving high-quality healthcare.

View full abstract

A 92-year-old woman was admitted to our hospital because of consciousness disturbance. Her conscious levels were III-300 and E1V1M1, according to the Japan Coma Scale and Glasgow Coma Scale, respectively. A blood test revealed a slightly increased level of inflammatory markers and markedly increased BUN and NH₃ levels. Her urine was dark brown and had precipitates and a fecal smell. Bacteriological culture of her urine showed Escherichia coli and Bacteroides fragilis. Electroencephalography showed slow waves with triphasic-like waves. Abdominal ultrasonography (US) showed localized mural thickening of the cystic bladder and linear or pointlike hyperechoic foci in the urinary bladder, suggesting the presence of air and a vesicointestinal fistula. Computed tomography using contrast medium injected through the urethra revealed saccular bulging of the bladder and leakage into the small intestine, indicating a vesicointestinal fistula. Her consciousness disturbance was considered to be caused by hyperammonemia due to the vesicointestinal fistula. Although vesicointestinal fistula is a rare disease leading to consciousness disturbance, US helps in the diagnosis of suspected cases.

View full abstract

We report on a physiological inspection system that, without the introduction of a departmental system, cooperates with the hospital information system (HIS). The National Hospital Organization Minami-Okayama Medical Center considered introducing a low-cost system rather than an expensive departmental system when we started using HIS. We constructed an inspection system that uses the hospital network to make the physiological inspection system cooperate with HIS. The physiological inspection system flow includes order, examination, and the report of accounts. Order and implementation use the HIS client, and examination uses a medical device. The pulmonary function test uses an order entry system, and other examinations use a non-order entry system. The examinations with the non-order entry system could allow us to browse waveforms and images by using the hospital network, and to make a report using the HIS. The physiological inspection system has a time series and statistics function. This system could have the same function as a departmental system, which was to reduce cost.

View full abstract

The detection or measurement of human heart-type fatty acid-binding protein (H-FABP) and troponin T is useful for the diagnosis of acute coronary syndrome. In this study, we compared the qualitative rapid H-FABP test kit Rapicheck H-FABP (Rapicheck; DS Pharma Biomedical Co., Ltd.) with the quantitative rapid H-FABP test kit Rapidchip H-FABP (Rapidchip; Sekisui Medical Co., Ltd.) and with a qualitative rapid test kit for troponin T (Trop T Sensitive; Roche Diagnostics K.K.). The characteristics of these kits were also determined. All 3 kits showed positive results in diseases such as acute myocardial infarction, angina pectoris, heart failure, aortic dissection, and takotsubo cardiomyopathy. When the results of the H-FABP and troponin T tests differed, patients were always positive for H-FABP and negative for troponin T. Some of the patients who were only positive for H-FABP were in the hyperacute phase of acute myocardial infarction, may have had elevation of H-FABP because of renal dysfunction, or else had skeletal muscle H-FABP in the blood. Two patients were negative for H-FABP using the Rapidchip test and positive using the Rapicheck test. The qualitative test is associated with the problem of individual variations due to subjective judgment. On the other hand, the Rapidchip allows the quantitative measurement of H-FABP and is free from this problem, making judgment of the results easy. Diagnostic sensitivity may be increased by using these kits in combination on the basis of the knowledge of the characteristics of each kit.

View full abstract

Regulations related to the work environment have been strengthened by the revisions of the Specific Chemical Disorder Prevention Rules (hereafter, the Rules, revised in March 2008) and the Regulations on Labor Standards for Women (hereafter, the Regulations, revised in October 2012). The standardization of the work environment with regard to its safety is demanded. In this study, we examined the realization of the pathological cytodiagnostic laboratories that complies with the Rules and the Regulations to improve the work environment. The following two work areas were prepared: 1) a work area that complies with the Rules and incorporates measures against infection and 2) a work area that complies with the Regulations. These two areas were not previously separated and used for sample preparation. Regarding 1), five local exhaust devices were installed, the pressure of a room was made negative to prevent the outflow of odor and harmful substances, the room temperature was set in the range of 22–25°C, and the room was equipped with a 24-hour ventilation system. In addition, a chemical closet with an exhaust device and a cabinet for formalin-fixed organ samples were installed with each equipped with a 24-hour ventilation system. A push-pull ventilation unit for formalin gas was placed in an organ-imaging device. Regarding 2), two existing Dalton draft chambers were used. The exhaust hoses of an automatic embedding device and an automatic staining device were connected to the exhaust duct of the Dalton draft chambers to obtain 24-hour ventilation. Owing to these measures, we were able to prepare controlled work areas of class I based on the work environment classification in compliance with the Rules and the Regulations. Raising the awareness and care of the staff involved alone in pathological examination are insufficient to achieve adequate measures. Obtaining the understanding of hospitals is important. Preparing a safe work environment and its complete management and control to prevent the diffusion of chemical substances and odor, in addition to the installation of suitable ventilation systems for the intended use, are required.

View full abstract

The Medical Technologist Working Group (WG) established in the Hyogo Prefectural Joint Committee of Transfusion Therapy (i.e., Committee) carried out some activities with the aim of improving safety in transfusion medicine in small- and medium-scale hospitals. The Medical Technologist WG performed practical skill training for blood transfusion inspection, as well as a joint workshop on blood transfusion manipulation with the Nurse WG. The annual questionnaire survey at the time of the practical skill training revealed that 18 out of 35 people attended in 2014 (51%), and 15 out of 30 people attended in 2015 (50%); they belonged to medical institutions with less than 200 beds. Twenty-one out of 35 people attended in 2014 (60%), and 17 out of 30 people attended in 2015 (57%); they did not exclusively engage in blood transfusion inspection duties in clinical laboratories. In fact, without specialists, it is considered difficult to acquire techniques and knowledge on blood transfusion inspection in clinical laboratories. To improve blood transfusion safety, it might be useful that practical skill training on blood transfusion inspection be held under the sponsorship of the Committee and the Hyogo Association of Medical Technologists.

View full abstract

Several factors may cause errors in the urinary sediment test, including centrifuge operation and supernatant removal. Previously, we reported a discrepancy between red blood cell (RBC) counts from flow cytometric analyses of noncentrifuged urine samples and urinary sediments after centrifugal separation, which was identified by basic evaluation at the entrance of the automated urine particle analyzer. To investigate the cause of this discrepancy, we examined RBCs remaining in the su2pernatant and RBC morphology after urinary sediment preparation. A significant amount of RBCs was observed to remain in the supernatant after centrifugal separation, leading to the above discrepancy. In addition, we found that RBCs that were judged “mixed” or “dysmorphic” by flow cytometric analysis were more likely to remain in the supernatant compared with RBCs that were judged “isomorphic”.

View full abstract

Laboratory tests are useful for improving diagnostic efficiency. Furthermore, they are essential for carrying out medical treatment. However, the environment surrounding the clinical laboratory is becoming increasingly severe, and laboratory testing is falling into a dangerous situation. Under these circumstances, unnecessary laboratory tests have been conducted without considering the prescribed limits. At our hospital, requests for laboratory tests are made using the electronic medical records. However, there have been occurrences of multiple tests exceeding the stipulated limits of the number of tests conducted on the same day for one patient, the number of tests conducted simultaneously, and the number of related tests. The introduction of a check function for each laboratory test that would display a warning when multiple tests are requested within a certain period of time would reduce the number of unnecessary laboratory tests.

View full abstract