Evaluation of lid speculum with a drape (LiDrape(®)) for preventing surgical-field contamination.

PURPOSE: To compare the degree of surgical-field contamination in cataract surgeries between a new draping method using a lid speculum with a drape (LiDrape(®)) and a conventional draping method. METHODS: Cataract surgery was performed on 21 eyes using LiDrape(®) (LiDrape(®) group) and on 22 eyes using a conventional draping method (conventional group). Contamination was evaluated by bacterial culture of conjunctival sac scrapings and ocular surface irrigation fluid. Conjunctival sac scrapings were collected before and after application of preoperative antibiotics. Ocular surface irrigation fluid was collected before incision placement and during surgery. Bacterial detection rate and types of organisms isolated at these four time points were examined. RESULTS: Bacterial detection rates were significantly decreased in the LiDrape(®) group at all time points after the application of antibiotics compared with preapplication. Regarding between-group comparisons, the bacterial detection rate in the LiDrape(®) group was only significantly lower than that in the conventional group in the intraoperative sample. Propionibacterium acnes was the most common organism isolated from ocular surface irrigation fluid. The number of P. acnes in the intraoperative sample was significantly lower in the LiDrape(®) group compared with the conventional group. There were no significant differences in detection rates for other bacteria between the groups. CONCLUSION: LiDrape(®) was as effective as conventional draping for preventing surgical-field contamination. The number of P. acnes during surgery was significantly lower in the LiDrape(®) group compared with the conventional group, suggesting that LiDrape(®) may contribute to the prevention of postoperative infection.

Online reporting system for transfusion-related adverse events to enhance recipient haemovigilance in Japan: a pilot study.

BACKGROUND: A surveillance system for transfusion-related adverse reactions and infectious diseases in Japan was started at a national level in 1993, but current reporting of events in recipients is performed on a voluntary basis. A reporting system which can collect information on all transfusion-related events in recipients is required in Japan. METHODS: We have developed an online reporting system for transfusion-related events and performed a pilot study in 12 hospitals from 2007 to 2010. RESULTS: The overall incidence of adverse events per transfusion bag was 1.47%. Platelet concentrates gave rise to statistically more adverse events (4.16%) than red blood cells (0.66%) and fresh-frozen plasma (0.93%). In addition, we found that the incidence of adverse events varied between hospitals according to their size and patient characteristics. CONCLUSION: This online reporting system is useful for collection and analysis of actual adverse events in recipients of blood transfusions and may contribute to enhancement of the existing surveillance system for recipients in Japan.

[A case of Desulfovibrio desulfuricans cultured from blood in Japan].

We report a case of Desulfovibrio desulfuricans bacteremia in a 60-year-old-man. In our case, anaerobic blood culture bottle turned out positive after five days' incubation. Gram stain showed the presence of slightly-curved Gram negative rod. Suspecting Campylobacter and Helicobacter, we added microaerobic culture while tentatively reporting Campylobacter to the physician. We then added anaerobic culturing with Brucella HK (RS) Agar because microaerobic culture proved the absence of microaerophile. We found small colonies on the third day, then we started anaerobic culture and eventually identified Desulfovibrio desulfuricans. We believe this is the first report of Desulfovibrio desulfuricans cultured from blood in Japan. In case Gram stain shows the presence of spiral bacterium, it is recommended to observe closely considering Desulfovibrio.

Rapid detection of haptoglobin gene deletion in alkaline-denatured blood by loop-mediated isothermal amplification reaction.

Anhaptoglobinemic patients run the risk of severe anaphylactic transfusion reaction because they produce serum haptoglobin antibodies. Being homozygous for the haptoglobin gene deletion allele (HP(del)) is the only known cause of congenital anhaptoglobinemia, and detection of HP(del) before transfusion is important to prevent anaphylactic shock. In this study, we developed a loop-mediated isothermal amplification (LAMP)-based screening for HP(del). Optimal primer sets and temperature for LAMP were selected for HP(del) and the 5' region of the HP using genomic DNA as a template. Then, the effects of diluent and boiling on LAMP amplification were examined using whole blood as a template. Blood samples diluted 1:100 with 50 mmol/L NaOH without boiling gave optimal results as well as those diluted 1:2 with water followed by boiling. The results from 100 blood samples were fully concordant with those obtained by real-time PCR methods. Detection of the HP(del) allele by LAMP using alkaline-denatured blood samples is rapid, simple, accurate, and cost effective, and is readily applicable in various clinical settings because this method requires only basic instruments. In addition, the simple preparation of blood samples using NaOH saves time and effort for various genetic tests.

Comparison of two control measures of weatherstripping in reducing blowing dust during hospital renovations.

Hospital renovation projects pose risks of invasive infection by fungi from dust that is blown about during the period in question. Control measures to reduce the amount of dust during hospital renovation are thus necessary. Currently, no study has compared different control measures for effectiveness through more than one period of renovation. In this study, we examined the capacities of two control measures of weatherstripping (0.15 mm poly film and adhesive tape) to reduce the amount of blowing dust during two different hospital renovations (in 2008 and 2009). The amount of dust in the air of the hospital before and during the renovation was measured about once a week in both 2008 and 2009, and the between-year and within-year differences were tested. Our study revealed that the weatherstripping used in 2009 (adhesive tape) was significantly more effective than the measures taken in 2008 (0.15 mm poly film) to reduce the amount of dust during the renovations (p < 0.001), while in both years the amount of dust became significantly higher during the renovations than before the renovations. Differences in the effectiveness of weatherstripping during renovations between floors of the hospital were not significant in both 2008 and 2009. The number of Aspergillus-positive samples did not significantly increase compared with the number observed before the start of the hospital renovations (2006-2007) in 2008 and 2009, respectively. The weatherstripping potentially reduced the associated risk of airborne fungal infection.

Development and validation of a SYBR Green I-based real-time polymerase chain reaction method for detection of haptoglobin gene deletion in clinical materials.

BACKGROUND: Anhaptoglobinemic patients run the risk of severe anaphylactic transfusion reaction because they produce serum haptoglobin (Hp) antibodies. Being homozygous for the Hp gene deletion (HP(del)) is the only known cause of congenital anhaptoglobinemia, and clinical diagnosis of HP(del) before transfusion is important to prevent anaphylactic shock. We recently developed a 5'-nuclease (TaqMan) real-time polymerase chain reaction (PCR) method. STUDY DESIGN AND METHODS: A SYBR Green I-based duplex real-time PCR assay using two forward primers and a common reverse primer followed by melting curve analysis was developed to determine HP(del) zygosity in a single tube. In addition, to obviate initial DNA extraction, we examined serially diluted blood samples as PCR templates. RESULTS: Allelic discrimination of HP(del) yielded optimal results at blood sample dilutions of 1:64 to 1:1024. The results from 2231 blood samples were fully concordant with those obtained by the TaqMan-based real-time PCR method. CONCLUSION: The detection rate of the HP(del) allele by the SYBR Green I-based method is comparable with that using the TaqMan-based method. This method is readily applicable due to its low initial cost and analyzability using economical real-time PCR machines and is suitable for high-throughput analysis as an alternative method for allelic discrimination of HP(del).

Usefulness of high-concentration calcium chloride solution for correction of activated partial thromboplastin time (APTT) in patients with high-hematocrit value.

INTRODUCTION: Pseudoprolongation of activated partial thromboplastin time (APTT) is a serious problem in anticoagulation therapy for patients with high hematocrit, such as cyanotic congenital heart diseases. APTT pseudoprolongation occurs when APTT assay is performed using routinely used vacuum sampling tubes containing citrate. Because the plasma fraction is small in high-hematocrit blood, the prescribed volume of citrate would be excessive for APTT assay, resulting in prolongation of clotting or APTT pseudoprolongation. CLSI--The Clinical and Laboratory Standards Institute (formerly NCCLS) method is the established method to correct the pseudoprolongation. However, the CLSI method needs repeated blood drawings and time-consuming, complicate procedures. Thus, alternative simple method is desired. METHOD: We examined whether APTT pseudoprolongation would be prevented by the increase in free calcium ion concentration by using high-concentration calcium chloride solution for the assay. Blood samples were obtained from 15 patients with high hematocrit (65+/-6%) who had cyanotic congenital heart disease. RESULT: Conventional APTT assay using 0.025 mol/L calcium chloride solution gave greater APTT compared with the CLSI method (51.7+/-11.8 vs. 34.6+/-4.7 s, p<0.001). However, when 0.035 mol/L calcium chloride solution was used, APTT (36.6+/-5.8 s) was similar to that obtained from the CLSI method. There was a good correlation in APTT values between high-calcium chloride solution method and the CLSI method (the slope=0.57, r2=0.49). CONCLUSION: High-calcium chloride solution method is useful to correct APTT pseudoprolongation. Because of the simplicity and the need of a single blood drawing, this method would reduce the burdens of not only patients but also clinical laboratory.

[Vibrio vulnificus pollution of imported frozen Black Tiger shrimps in Japan].

The Ariake Sea area of Japan is endemic for Vibrio vulnificus infection. V vulnificus was isolated from slime from tidal flats, seawater, and fish Sea year-round as we reported previously. To identify new routes and factors of V vulnificus infection, we studied V. vulnificus pollution of imported frozen Black Tiger shrimps purchased from a fish market in Kurume, Fukuoka, Japan. V. vulnificus was isolated from 9 of 100 tails (9%) of Philippines products, 3 of 100 tails (3%) of Indonesia products, and 0 out of 100 tails (0%) of Madagascar products. Cytotoxin-hemolysin genes were identified in 7 V. vulnificus strains isolated from patients with V vulnificus septicemia, 9 strains from Philippine products, and 3 strains from Indonesian products. These results suggest that imported frozen Black Tiger shrimps are a new sources of V. vulnificus infection.

[Antibody titers against measles, rubella, mumps and varicella-zoster viruses in medical students].

It is important to identify and immunize susceptible students who have clinical practice to prevent and control hospital infections. The antibody titers to measles, rubella, mumps and varicella viruses were measured in 1,139 students(417 men, 722 women, average age 21.3+/-2.7 yr old)including 510 medical students, 442 nursing students and 187 students of the School of Medical Technology in Kurume University. Antibodies against measles virus were detected by particle agglutination assay(PA), those against rubella virus by hemagglutination inhibition assay(HI), and those against mumps and varicella viruses by enzyme-linked immunosorbent assay(EIA). The serological susceptibilities to measles, rubella, mumps and varicella viruses were 112(9.8%), 112(9.8%), 163(14.3%)and 73(6.4%), respectively. The serological susceptibilities to measles, rubella and mumps viruses in male students were not different from those in female students. The susceptibility to varicella virus in female students was significantly higher than that in male students. After susceptible students were recommended to have vaccinations against each virus, the vaccination rate of the students without antibody was 99.1%. The history of infection and vaccination against the viruses were examined by self-recorded questionnaires in 406 students from all disciplines. The serological susceptibility of students with positive vaccination history was 11.1% for measles, 6.8% for rubella, 18.3% for mumps, and 4.9% for varicella. The serological susceptibility of students with a positive infection history was 5.7% for measles, 3.4% for rubella, 2.9% for mumps, and 4.9% for varicella. In the self-recorded questionnaire, the rate of unknown infection and vaccination histories were 57.5% and 71.6% for measles, 52.5% and 68.4% for rubella, 34.3% and 75.6% for mumps, and 27.1% and 80.5% for varicella, respectively. In conclusion, these data confirm that it is essential to assess immune status against measles, rubella, mumps and varicella in students who have clinical practice in hospital regardless of infection or vaccination history. Accordingly, susceptible students should be vaccinated to prevent those viral infections in hospital.

Characteristic features of QRST integral mapping in patients with high risk Brugada syndrome.

BACKGROUND: The characteristic features of QRST integral mapping in the Brugada-type resting ECG of patients at a high risk for life-threatening ventricular arrhythmias were examined. METHODS AND RESULTS: QRST integral mapping was performed in 11 Brugada-type ECG patients with histories of aborted sudden death, spontaneous ventricular tachycardia and fibrillation (VT/VF) or programmed electric stimulation-inducible VT/VF (high risk group); 13 Brugada-type ECG patients without a history of such events (low risk group); and 21 age-matched healthy controls. Individual QRST isointegral maps revealed the minimum integral in the mid-to-right upper chest in 100% and 85% of the control and low risk groups, respectively, whereas this integral was 64% in the upper right back of the high risk group (p<0.05). On the QRST integral departure maps, the abnormal positive departure area (integral value>or=+2 standard deviation) was located in the mid-to-right upper chest in 82% and 8% of the high and low risk groups, respectively (p<0.05). During the follow-up period, sudden death or VF occurred in 4 of 6 high risk patients with both the abnormal findings. CONCLUSION: The abnormal positive departure area in the mid-to-right upper chest and the minimum QRST integral in the right upper back were distinct hallmarks for screening patients with the high risk Brugada-type ECG.

[Guidelines for safer and more appropriate use of blood transfusion systems in Japan].

The Blood Transfusion Law has been newly established in Japan since 2003. Under the new law, physicians and co-medical staff have to work hard to establish safer blood transfusion systems and more appropriate transfusion of blood components for the patients in their hospitals. Guide lines for transfusion of blood components made by the Ministry of Health, Welfare, and Labor in Japan are useful tools to accomplish the aims shown above. When they would transfuse blood components appropriately to the patients according to the guidelines, more fresh frozen plasma could be available for the raw materials to produce albumin components and immunoglobulin components those are now partially imported from the foreign countries. Eventually, blood components consumed in Japan for transfusion will be supplied by 100% inland donors blood.

[Antifungal susceptibility of Candida species causing candidemia in Kurume University Hospital and Shaiseikai Hutukaichi Hospital].

Prevention of candidemia has been difficult and empirical therapy may eventually reduce morbidity and mortality. Successful empirical therapies depend on understanding of fungal features and antifungal agents. Susceptibility to amphotericin B (AMPH-B), flucytosine (5-FC), fluconazole (FLCZ), itraconazole (ITCZ), miconazole (MCZ), and micafungin (MCFG) of 41 Candida species isolated from blood were determined. Candida albicans was the most common species (23 species), followed by C. parapsilosis (5 species), C. tropicalis (4 species), C. glabrata (3 species), C. guilliermondii (2 species), C. krusei (1 specie), and Candida spp (3 species). The isolation rates of the drug-resistant (DR) fungi were 5% for 5-FC. The rates of DR and susceptible dose dependent (S-DD) fungi were 0% and 2% for FLCZ, respectively. The rates of DR and S-DD fungi were 0% and 17% for ITCZ, respectively. No shift to resistant species in C. albicans occurred in our hospitals. All C. albicans were susceptible for the antifungal agents examined.

[Seasonal change of Vibrio vulnificus in the slime of tidal flats, seawater, and fishes collected along Ariake Sea, Japan].

To investigate the seasonal change of Vibrio vulnificus in Ariake Sea, Japan, we attempted to isolate V. vulnificus from the slime of tidal flats, seawater, and fishes obtained from three harbors along Ariake Sea. The sample were collected twice a month from January to December, 2001. Also, we determined the biological characteristics of the individual isolates. V. vulnificus were isolated throughout the year, but the isolation ratios were higher during the warmer season from June to October. The isolates in the warmer season were able to grow in culture media containing 0.5% NaCl, whereas those isolated in the remaining seasons could not. Moreover, the isolates in the warmer season showed a greater hemolytic activity than those isolated in the remaining seasons. These results suggest that V. vulnificus isolated in warmer seasons are more vigorous in nature than those isolated in the remaining seasons.

Heterozygosity for two novel null alleles of the KEL gene causes the Kell-null phenotype in a Japanese woman.

The Kell-null (Ko) phenotype is rare and it does not express the Kell antigens on erythrocyte membranes. Recently, several distinct missense and nonsense base substitutions in the coding region and the donor splice site of intron 3 were identified in the KEL gene in individuals with the Ko phenotype. We analysed both genomic DNA and cDNA sequences of the KEL gene in a Japanese woman with the Ko phenotype. She was found to be heterozygous for two novel null KEL alleles. One allele contained an A to G substitution in intron 5 that changes the 3'-splice site of intron 5 from AAG to AGG, resulting in a reading frameshift by a single guanine insertion in KEL mRNA, and the other allele contained a single G to A substitution in exon 12 (codon 459) creating a termination codon. Neither mutation was found in 114 randomly selected Japanese individuals. The results suggested that the Ko blood group phenotype might be owing to several distinct non-functional alleles without any prevalent allele.

[Drug sensitivity and beta-lactamase producibility of various types of bacteria clinically isolated during the period from December 1999 to February 2000].

beta-Lactamase activity and drug sensitivity were measured in 744 strains from 8 species of bacteria isolated at medical institutions in Chikugo District of Fukuoka Prefecture during the period from December 1999 to February 2000. Nitrocefin test revealed that beta-lactamase was positive in 48% of S. aureus, 7% of H. influenzae, and 92% of M. catarrhalis, and acidometry revealed that penicillinase/cephalosporinase were positive in 13%/14% of E. coli, 22%/8% of K. pneumoniae, 47%/97% of E. cloacae, 3%/65% of S. marcescens, and 10%/36% of P. aeruginosa. Based on the assessment of the MIC values of various types of antibacterial drugs for beta-lactamase-producing strains, there were 11 strains (1 strain of K. pneumonia, 6 strains of E. cloacae, and 4 strains of P. aeruginosa) of class-B beta-lactamase-producing bacteria out of a total of 496 strains of Enterobacteriaceae and Pseudomonas aeruginosa. The results of PCR analysis suggested that 1 strain of K. pneumonia, 1 strain of E. cloacae, and 4 strains of P. aeruginosa produced metallo-beta-lactamase. There was no strain (E. coli and K. pneumoniae) of ESBL-producing bacteria. BLNAR strains, on the other hand, were found in 9% (9/100) of H. influenzae.