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ABSTRACT: Hybrisep is an in situ hybridization (ISH) method to detect phagocytosed bacteria in peripheral blood neutrophils and macrophages. We report 10 actual clinical cases tested using Hybrisep with new DNA probes, and the data were compared to the actual blood culture results. A normal Hybrisep strategy employs 5 DNA probes to detect the following bacterial DNA: "SA" probe for S. aureus, "SE" for S. epidermidis, "PA" for P. aeruginosa, "EF" for E. faecalis, and "EK" for E. coli, E. cloacae, and K. pneumoniae. Six newly designed DNA probes were used in this study: "GB" probe for 49 common bacteria, "SP" for S. pneumoniae, "BF" for B. fragilis, "HI" for H. influenzae, "GC" for Candida species, and "CA" for C. albicans. Three cases were positive on ISH, but all their blood cultures were negative. One case showed a positive blood culture, but was negative on ISH. In another 6 cases, both were negative. We postulated that empirical therapy of antibiotics resulted in only positive ISH. Cases only showing a positive outcome on blood culture might be due to a diminished phagocytic function during patients' severe disease conditions. In conclusion, ISH with Hybrisep has clinical advantages such as being able to defect causative pathogens even after the use of antibiotics, and facilitates more rapid identification than routinely performed bacterial cultures only.
Rinsho byori. The Japanese journal of clinical pathology 12/2010; 58(12):1221-9.
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Miho Okutsu,
Hitoshi Ohto,
Hiroyasu Yasuda,
Kinuyo Kawabata,
Satoshi Ono,
Shunnichi Saito,
Akiko Sugawara,
Masami Kikuchi, Saori Miura,
Youko Ishii,
Kazuya Watanabe,
Yuriko Tohyama,
Kenneth E Nollet
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ABSTRACT: The indirect antiglobulin test (IAT) can be potentiated by agents such as polyethylene glycol (PEG-IAT) and albumin (Alb-IAT). PEG-IAT is generally regarded as superior to Alb-IAT for the detection of clinically significant red blood cell (RBC) antibodies. However, supporting data come from Caucasian-dominant populations. Non-Caucasian populations should be investigated as well.
In this single-centre, retrospective, sequential study, Alb-IAT was used from 1989 to 1996 (8 years) and PEG-IAT from 1997 to 2008 (12 years). Pre-transfusion RBC alloantibody detection rates and specificity, post-transfusion alloantibody production, and the incidence of delayed haemolytic transfusion reaction were assessed and compared for the two periods.
Although overall RBC alloantibody detection rates were comparable, PEG-IAT more frequently detected clinically significant antibodies such as anti-E, anti-Fy(b), and anti-Jk(a), and less frequently detected insignificant antibodies such as anti-Le(b) and anti-P(1). New alloantibodies emerged comparably during the two periods. Delayed haemolytic transfusion reaction was less frequent during the PEG-IAT period (0.30% versus 0.12%, p<0.05).
PEG-IAT was superior in the detection of clinically significant antibodies, reduced the detection of insignificant antibodies, and prevented delayed haemolytic transfusion reaction better than Alb-IAT among Japanese transfusion recipients in this retrospective survey of limited power.
Blood transfusion = Trasfusione del sangue 11/2010; 9(3):311-9. · 2.10 Impact Factor
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Pediatrics International 09/2009; 51(4):585-7. · 0.63 Impact Factor
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Hiroyasu Yasuda,
Miho Okutsu,
Kinuyo Kawabata, Saori Miura,
Chikako Baba,
Satoshi Ono,
Yoko Gunji,
Kazuya Watanabe,
Yuko Obata,
Ryoko Kikuchi,
Hitoshi Ohto
Japanese Journal of Transfusion and Cell Therapy 01/2007; 53(6):613-618.
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ABSTRACT: The severity of hemolytic disease of the newborn (HDN) due to Diego(b) (Di(b)) mismatch ranges from no symptoms to severe jaundice that requires exchange transfusion (ET). The clinical significance of anti-Di(b) is incompletely recognized.
A male newborn, referred with jaundice, was revealed to have HDN due to Di(b) mismatch and was treated successfully with phototherapy and high-dose intravenous gamma globulin (IVGG).
The literature of HDN caused by Di(b) mismatch was reviewed. The cases were classified into three groups according to their severity: the mildest needed no therapy (NO), the moderate group received phototherapy alone (PHOTO), and the most severe was treated with ET and/or high-dose IVGG therapy plus phototherapy (ET/IVGG).
Among 27 cases of HDN due to Di(b) reported to date, 10, 6, and 11 cases required NO, PHOTO, and ET/IVGG, respectively. A significant correlation (p < 0.01) was found between the maternal anti-Di(b) titer and the severity of the disease when the ET/IVGG group was compared with the NO group. All mothers of the group that needed ET/IVGG had an anti-Di(b) titer of 64 or greater.
A maternal high titer (> or =64) of anti-Di(b) is associated with a higher risk of severe hyperbilirubinemia for mismatched newborns.
Transfusion 04/2006; 46(3):454-60. · 3.22 Impact Factor
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ABSTRACT: BACKGROUND: The severity of hemolytic disease of the newborn (HDN) due to Diegob (Dib) mismatch ranges from no symptoms to severe jaundice that requires exchange transfusion (ET). The clinical significance of anti-Dib is incompletely recognized.CASE REPORT: A male newborn, referred with jaundice, was revealed to have HDN due to Dib mismatch and was treated successfully with phototherapy and high-dose intravenous gamma globulin (IVGG).STUDY DESIGN AND METHODS: The literature of HDN caused by Dib mismatch was reviewed. The cases were classified into three groups according to their severity: the mildest needed no therapy (NO), the moderate group received phototherapy alone (PHOTO), and the most severe was treated with ET and/or high-dose IVGG therapy plus phototherapy (ET/IVGG).RESULTS: Among 27 cases of HDN due to Dib reported to date, 10, 6, and 11 cases required NO, PHOTO, and ET/IVGG, respectively. A significant correlation (p < 0.01) was found between the maternal anti-Dib titer and the severity of the disease when the ET/IVGG group was compared with the NO group. All mothers of the group that needed ET/IVGG had an anti-Dib titer of 64 or greater.CONCLUSION: A maternal high titer (≥64) of anti-Dib is associated with a higher risk of severe hyperbilirubinemia for mismatched newborns.
Transfusion 02/2006; 46(3):454 - 460. · 3.22 Impact Factor
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ABSTRACT: It has been controversial whether HLA antibodies cause hemolytic transfusion reactions (HTR) or shortened red blood cell (RBC) survival. A patient is reported who had two episodes of HTR, the latter of which was likely due to RBC-reactive HLA antibodies.
A 77-year-old woman, admitted for gastric varix rupture, had no RBC-irregular antibodies detected before transfusion. On Hospital Day 12, after transfusion of 2 units of RBCs and 2 units of fresh-frozen plasma, the first delayed hemolytic episode occurred and anti-E, anti-c, anti-Jk(a), and unidentified RBC-reactive antibodies were detected in a serum sample from Day 14. Two additional units of matched RBCs were transfused with a leukoreduction filter on Days 19 and 22. After 4 hours of starting a transfusion on Day 22, the patient had fever, and a second hemolytic episode was recorded. Multireactive HLA antibodies (reactive against 20 of 20 donor panel lymphocytes) were detected in serum samples from Day 15 to Day 21. These HLA antibodies reacted strongly with HLA-A2 and HLA-B7 antigens, corresponding to Bg(c) and Bg(a) antigens on RBCs, respectively. RBCs transfused on Day 22 were found to be HLA-A2 by genotyping.
Strong HLA alloantibodies in this recipient appear to have caused a HTR. It is suggested that HLA antibodies be considered in patients with unexplained HTRs.
Transfusion 01/2006; 45(12):1925-9. · 3.22 Impact Factor
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Chikako Takeuchi,
Hitoshi Ohto, Saori Miura,
Hiroyasu Yasuda,
Satoshi Ono,
Takashi Ogata,
千華子 竹内,
斉 大戸,
里織 三浦,
広康 安田,
智 小野,
隆 尾形
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ABSTRACT: Background：It has been controversial whether HLA antibodies cause hemolytic transfusion reactions (HTR) or shortened RBC survival. We report a patient who had two episodes of HTR, the latter of which was likely due to RBC-reactive HLA antibodies. Case Report：A 77-year-old woman, admitted for gastric varix rupture, had no RBC irregular antibodies detected before transfusion. On hospital day 12, after transfusion of two units of RBCs and two units of FFP, the first delayed hemolytic episode occurred and anti-E, anti-c, anti-Jka and unidentified RBC-reactive antibodies were detected in serum from day 14. Two further units of compatible RBCs were transfused using a leukocyte-reduction filter on days 19 and 22. After 4 hours of starting a transfusion on day 22, the patient had fever, and a second hemolytic episode was recorded. Multireactive HLA antibodies (reactive against 20 of 20 donor panel lymphocytes) were detected in sera from day 15 to day 21. These HLA antibodies reacted strongly with HLA-A2 and HLA-B7 antigens, corresponding to Bgc and Bga antigens on RBCs, respectively. RBCs transfused on day 22 were found to be HLA-A2 by genotyping.
Conclusion： Strong HLA alloantibodies in this recipient appear to have caused a HTR. It is suggested that HLA antibodies be considered in patients with unexplained HTRs.
