ABSTRACT
We collected data from all the women with singleton pregnancies complicated by early-onset severe preeclampsia between 2008 and 2018 to identify the factor(s) that contributed to favourable neonatal outcome. Thirty women delivered the neonates with favourable outcome and the remaining 21 women delivered those with unfavourable outcome. Univariate logistic regression analysis revealed that gestational age at diagnosis ≥28 weeks (crude odds ratio [OR], 6.00), protocol-based management (crude OR 5.83), use of magnesium sulphate (crude OR, 6.00), gestational age at delivery ≥32 weeks (crude OR, 31.90), and birthweight ≥1000 g (crude OR, 10.36) were significantly associated with favourable neonatal outcome. Among these five factors, multivariate logistic regression analysis extracted gestational age at delivery ≥32 weeks (adjusted OR, 17.62) as an only independent factor contributing to favourable neonatal outcome. These data suggest that prolongation of pregnancy up to 32 weeks of gestation is a key factor to improve neonatal outcome in the expectant management of early-onset severe preeclampsia.This study was approved by the ethics committee of Otsu Red Cross Hospital (reference number: 363, date of approval: April 28th, 2016).Impact statementWhat is already known on this subject? It has been demonstrated that expectant management for the women with early-onset severe preeclampsia is associated with decreased neonatal morbidity as compared to the aggressive management, suggesting that prolongation of the pregnancy period contributes to improved neonatal outcomes.What do the results of this study add? Among multiple elements composing expectant management for the women with early-onset severe preeclampsia, 'gestational age at delivery ≥32 weeks' was extracted as an only independent factor that significantly contributes to favourable neonatal outcomes. Importantly, small for gestational age was not significantly associated with poor neonatal outcomes.What are the implications of these findings for clinical practice and/or further research? The obstetrician should aim to prolong the pregnancies complicated by early-onset severe preeclampsia up to 32 gestational weeks even in the presence of foetal growth restriction, as far as maternal conditions allow. Such management policy could contribute to improvement of the neonatal outcomes.
Subject(s)
Gestational Age , Pre-Eclampsia/physiopathology , Pregnancy Outcome/epidemiology , Premature Birth/prevention & control , Adult , Birth Weight , Clinical Protocols , Female , Fetal Growth Retardation , Humans , Infant, Newborn , Logistic Models , Magnesium Sulfate/therapeutic use , Odds Ratio , Pre-Eclampsia/therapy , Pregnancy , Pregnancy Trimester, Third , Premature Birth/etiology , Treatment OutcomeABSTRACT
BACKGROUND: The reliable diagnosis of human T-cell leukemia virus type 1 (HTLV-1) infection is important, particularly as it can be vertically transmitted by breast feeding mothers to their infants. However, current diagnosis in Japan requires a confirmatory western blot (WB) test after screening/primary testing for HTLV-1 antibodies, but this test often gives indeterminate results. Thus, this collaborative study evaluated the reliability of diagnostic assays for HTLV-1 infection, including a WB-based one, along with line immunoassay (LIA) as an alternative to WB for confirmatory testing. RESULTS: Using peripheral blood samples from blood donors and pregnant women previously serologically screened and subjected to WB analysis, we analyzed the performances of 10 HTLV-1 antibody assay kits commercially available in Japan. No marked differences in the performances of eight of the screening kits were apparent. However, LIA determined most of the WB-indeterminate samples to be conclusively positive or negative (an 88.0% detection rate). When we also compared the sensitivity to HTLV-1 envelope gp21 with that of other antigens by LIA, the sensitivity to gp21 was the strongest. When we also compared the sensitivity to envelope gp46 by LIA with that of WB, LIA showed stronger sensitivity to gp46 than WB did. These findings indicate that LIA is an alternative confirmatory test to WB analysis without gp21. Therefore, we established a novel diagnostic test algorithm for HTLV-1 infection in Japan, including both the performance of a confirmatory test where LIA replaced WB on primary test-reactive samples and an additional decision based on a standardized nucleic acid detection step (polymerase chain reaction, PCR) on the confirmatory test-indeterminate samples. The final assessment of the clinical usefulness of this algorithm involved performing WB analysis, LIA, and/or PCR in parallel for confirmatory testing of known reactive samples serologically screened at clinical laboratories. Consequently, LIA followed by PCR (LIA/PCR), but neither WB/PCR nor PCR/LIA, was found to be the most reliable diagnostic algorithm. CONCLUSIONS: Because the above results show that our novel algorithm is clinically useful, we propose that it is recommended for solving the aforementioned WB-associated reliability issues and for providing a more rapid and precise diagnosis of HTLV-1 infection.
Subject(s)
Algorithms , Diagnostic Tests, Routine/methods , HTLV-I Infections/diagnosis , Human T-lymphotropic virus 1/isolation & purification , Antibodies, Viral/blood , Blotting, Western , Diagnostic Tests, Routine/standards , HTLV-I Antigens/immunology , Human T-lymphotropic virus 1/immunology , Humans , Immunoassay , Japan , Polymerase Chain Reaction , Proviruses/genetics , Proviruses/isolation & purification , Reagent Kits, Diagnostic , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
BACKGROUND: In 2014, we experienced the first isolation of Lactococcus garvieae from a platelet concentrate (PC). Thereafter, L. garvieae contamination of PCs occurred in two more cases in Japan. It is rare that bacterial contamination with uncommon strains like this species occurs frequently within a short period. Therefore, we performed a detailed analysis of the characteristics of these strains. STUDY DESIGN AND METHODS: Three bacterial strains were identified by biochemical testing and molecular analysis. Genomic diversity was characterized by multilocus sequence typing (MLST). To observe growth kinetics in blood components, PCs were inoculated with the three different strains. RESULTS: All three strains were identified as L. garvieae by molecular analysis. Each strain belonged to a different phylogenetic group according to MLST analysis. In the spiking trial, the three strains demonstrated differences in their final concentrations and changes in appearance of PCs. CONCLUSION: In this study, all three L. garvieae strains were correctly identified by molecular analysis. Since the three strains were collected in different regions of Japan and belonged to different phylogenetic groups according to MLST analysis, it is suggested that L. garvieae have a wide distribution with diversity in Japan. In PCs, the three L. garvieae strains showed clear differences in growth kinetics and changes in appearance of PCs. These differences may have been the primary determinant of whether PC contamination was detected before transfusion. Moreover, L. garvieae represents an emerging foodborne bacterium that can cause transfusion-transmitted bacteremia. Understanding our cases may help prevent bacterial contamination of blood products.
Subject(s)
Blood Platelets/microbiology , Lactococcus , Phylogeny , Humans , Lactococcus/classification , Lactococcus/genetics , Lactococcus/growth & development , Lactococcus/isolation & purification , Multilocus Sequence TypingABSTRACT
BACKGROUND: Chagas disease is caused by Trypanosoma cruzi and is endemic in Latin America. In nonendemic countries, including Japan, Chagas disease is primarily a problem in the context of transfusion transmission. Approximately 250,000 immigrants from Latin America reside in Japan, and many of those individuals serve as active blood donors. This study surveyed the seroprevalence of T. cruzi infection among at-risk blood donors in Japan, defined as those who themselves (or whose mothers) were born (or raised) in Latin America, or those with a travel history to Latin America. STUDY DESIGN AND METHODS: Blood samples were obtained from at-risk donors in two periods, 2004-2012 and 2013-2016. Collected samples were tested for T. cruzi antibodies using both an enzyme-linked immunosorbent assay and a chemiluminescent immunoassay. Samples that tested positive in both assays were additionally tested by polymerase chain reaction, and look-back investigation was conducted when necessary. RESULTS: Of 18,484 samples obtained from 18,076 at-risk donors, 3 (1:6,025, 0.017%) donors showed seroreactivity by enzyme-linked immunosorbent assay and chemiluminescent immunoassay. All antibody-positive donors were born in Latin America. One of them also was positive for T. cruzi DNA. Eleven previous donations from this donor were subjected to look-back investigation, and five recipients were tested. All five recipients tested negative for T. cruzi antibodies. CONCLUSION: Seroprevalence of T. cruzi was 0.017% among at-risk donors in Japan. Transfusion-transmitted infection of Chagas disease has not been confirmed to date. Screening for T. cruzi antibodies by targeting at-risk donors is an appropriate strategy for ensuring blood safety in Japan.
Subject(s)
Chagas Disease/epidemiology , Trypanosoma cruzi/pathogenicity , Antibodies, Protozoan/immunology , Blood Donors/statistics & numerical data , Chagas Disease/immunology , DNA, Protozoan/genetics , Female , Humans , Immunoassay , Japan , Male , Trypanosoma cruzi/geneticsABSTRACT
Quantitative PCR (qPCR) of human T-cell leukemia virus type 1 (HTLV-1) provirus is used for HTLV-1 testing and for assessment of risk of HTLV-1-related diseases. In this study, a reference material was developed for standardizing HTLV-1 qPCR. Freeze-dried TL-Om1 cells diluted with Jurkat cells were prepared and an assigned value for proviral load (PVL) of 2.71 copies/100 cells was determined by digital PCR. Nine Japanese laboratories using their own methods evaluated the PVLs of this reference material as 1.08-3.49 copies/100 cells. The maximum difference between laboratories was 3.2-fold. Correcting measured PVLs by using a formula incorporating the assigned value of this reference material should minimize such discrepancies.
Subject(s)
DNA, Viral/analysis , Human T-lymphotropic virus 1/genetics , Leukemia, T-Cell/virology , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/standards , Cell Line, Tumor , DNA, Viral/genetics , Disaccharides/genetics , HTLV-I Infections/genetics , HTLV-I Infections/virology , Humans , Japan , Jurkat Cells , Proviruses/genetics , Reference Standards , Viral Load/geneticsABSTRACT
There are few reports on HIV-1 intra-host evolutionary rate in asymptomatic treatment-naïve patients. Here, the HIV-1 intra-host evolutionary rate was estimated based on HIV-1 RNA sequences from plasma samples of blood donors in Japan. Blood donors were assumed to have received no treatment for and have no symptoms of HIV-1 infection because they were healthy, and declared no risky behaviors of HIV-1 infection on a self-reported questionnaire or interview followed by donation. HIV-1 RNA was obtained from 85 plasma samples from 36 blood donors who donated blood multiple times and were HIV-1-positive. The C2V3C3 region which encodes for a part of the envelope protein, and the V3 loop in the C2V3C3 region were analyzed by RT-PCR and direct sequencing, and the sequences were compared. The nucleotide substitution rate was calculated by linear regression. All HIV-1 samples analyzed were classified as subtype B. The mean nucleotide substitution rate in C2V3C3 was calculated to be 6.2 × 10-3-1.8 × 10-2/site/year (V3: 4.5 × 10-3-2.3 × 10-2/site/year). The mean non-synonymous substitution rate in C2V3C3 was calculated to be 5.2 × 10-3-1.7 × 10-2/site/year (V3: 4.5 × 10-3-2.1 × 10-2/site/year). The mean synonymous substitution rate in C2V3C3 was calculated to be 1.1 × 10-4-2.3 × 10-3/site/year (V3: 2.9 × 10-3/site/year). Among HIV-1 subtype B RNA-positive blood donors in Japan, the nucleotide substitution rate in C2V3C3 was estimated to be higher than that of reported cases using HIV-1 samples mainly obtained from AIDS patients. Compared to AIDS patients, immune responses against HIV-1 are probably more effective in HIV-1 RNA-positive blood donors. Consequently, immune pressure presumably promotes mutation of the virus genome.
Subject(s)
Evolution, Molecular , HIV-1/genetics , Mutation Rate , Blood Donors , HIV Infections/virology , HIV-1/classification , Humans , Japan , Male , RNA, Viral , Sequence Analysis, RNAABSTRACT
BACKGROUND: Human babesiosis is caused mainly by Babesia microti and has recently become a public health concern due to an increase in transfusion-transmitted infection. Thus, the development of an antibody detection method with high specificity and sensitivity is a priority. Seroreactivity against B. microti has been reported to be highly specific not only to B. microti lineages but also to sublineages. This study aimed to elucidate the human antibody reactivity against various lineages, including US, Kobe, and Hobetsu, and sublineages (North America and East Asia) in the US lineage. STUDY DESIGN AND METHODS: Twenty samples obtained from individuals infected with B. microti in the United States were tested for the presence of anti-B. microti antibodies using indirect immunofluorescence assay (IFA) and Western blotting (WB) to indicate antigens of each (sub-)lineage. RESULTS: By IFA, 20 samples showed reactivity to the North America sublineage (titer range, 64-4096), 16 to the East Asia sublineage (64-512), 10 to the Kobe (64-128), and five to the Hobetsu (64). Antibody titers to the East Asia sublineage, Kobe, and Hobetsu were significantly lower than those to the North America sublineage (p < 0.01). By WB, in parallel with the IFA results, 18 samples showed strong reactions to the North America sublineage, weak reactions to the East Asia sublineage, and near-zero reactions to the Kobe and Hobetsu. CONCLUSION: Human antibodies induced by B. microti infection are highly specific against B. microti lineages and sublineages with low cross-reactivity. Developing a precise antibody detection method may require specific antigens based on B. microti lineages and sublineages.
Subject(s)
Babesia microti/immunology , Babesiosis/diagnosis , Cross Reactions/immunology , Animals , Antibodies, Protozoan , Antigens, Protozoan , Humans , North America , Parasites/immunologyABSTRACT
Western blotting (WB) for human T cell leukemia virus type 1 (HTLV-1) is performed to confirm anti-HTLV-1 antibodies detected at the initial screening of blood donors and in pregnant women. However, the frequent occurrence of indeterminate results is a problem with this test. We therefore assessed the cause of indeterminate WB results by analyzing HTLV-1 provirus genomic sequences. A quantitative PCR assay measuring HTLV-1 provirus in WB-indeterminate samples revealed that the median proviral load was approximately 100-fold lower than that of WB-positive samples (0.01 versus 0.71 copy/100 cells). Phylogenic analysis of the complete HTLV-1 genomes of WB-indeterminate samples did not identify any specific phylogenetic groups. When we analyzed the nucleotide changes in 19 HTLV-1 isolates from WB-indeterminate samples, we identified 135 single nucleotide substitutions, composed of four types, G to A (29%), C to T (19%), T to C (19%), and A to G (16%). In the most frequent G-to-A substitution, 64% occurred at GG dinucleotides, indicating that APOBEC3G is responsible for mutagenesis in WB-indeterminate samples. Moreover, interestingly, five WB-indeterminate isolates had nonsense mutations in Pol and/or Tax, Env, p12, and p30. These findings suggest that WB-indeterminate carriers have low production of viral antigens because of a combination of a low proviral load and mutations in the provirus, which may interfere with host recognition of HTLV-1 antigens.
Subject(s)
Antibodies, Viral/immunology , HTLV-I Infections/diagnosis , Human T-lymphotropic virus 1/genetics , Human T-lymphotropic virus 1/immunology , Proviruses/genetics , APOBEC-3G Deaminase/metabolism , Blood Donors , Blotting, Western , Cell Line , Codon, Nonsense/genetics , Female , Genome, Viral/genetics , HTLV-I Infections/virology , Humans , Pregnancy , Real-Time Polymerase Chain Reaction/methods , Serologic Tests/methods , Viral Load , Virus Replication/geneticsABSTRACT
Adult T-cell leukemia/lymphoma (ATL) occurs in approximately 5% of individuals infected with human T-cell leukemia virus type 1 (HTLV-1). A high proviral load (PVL; more than four copies per 100 peripheral blood mononuclear cells (PBMCs) or 1.6 copies per 100 blood leukocytes) and being male are risk factors for ATL development. Whether anti-HTLV-1 antibody level is related to such risk is unknown. Here, PVL and antibody levels were examined using real-time PCR and other tests in 600 HTLV-1 positive screened Japanese blood donors to understand the relationship between PVL and antibody level in asymptomatic carriers and to gain insights toward better antibody testing for HTLV-1 infection. The 430 donors in whom proviral DNA was detected were considered as true positives for HTLV-1 infection. Among donors aged 40 years or older, more males than females had a PVL corresponding to more than 1.6% infected leukocytes, and an antibody titer below the median (P = 0.0018). In antibody tests using an HTLV-1 positive cell line or Env antigens there was a large discrepancy in antibody titer among 13 provirus-positive samples, probably suggesting that antibody-based screening tests should incorporate multiple HTLV-1 antigens, such as Gag and Env antigens.
Subject(s)
Antibodies, Viral/blood , Blood Donors , HTLV-I Infections/immunology , HTLV-I Infections/virology , Human T-lymphotropic virus 1/isolation & purification , Proviruses/isolation & purification , Viral Load , Adolescent , Adult , Aged , Carrier State/immunology , Carrier State/virology , Female , Humans , Japan , Male , Middle Aged , Young AdultABSTRACT
In Japan, the number of human immunodeficiency virus (HIV)-1 infections remains relatively low; nevertheless, the annual incidence of HIV-1 infection has not decreased. New infections remain a great concern, and an improved understanding of epidemiological trends is critical for public health. The env C2V3 and pol sequences of HIV-1 RNA from 240 early (1996-2001) and 223 more recent (2010-2012) blood donations were used to compare the distribution of virus subtypes and to generate phylogenetic trees. Subtype B was clearly predominant in both early and more recent donations (both were 88.3%), and CRF01_AE was the second most common subtype. Phylogenetic analysis revealed a peculiar epidemiological transition. Compared to early subtype B isolates from 2 major endemic areas (Tokyo and Osaka), the more recent subtype B isolates formed fewer tight clusters in phylogenetic trees (from 8 to 2 clusters in Tokyo and 5 to zero clusters in Osaka). Furthermore, mixing of HIV-1 infections between these 2 endemic areas appear to increase. Analysis of phylogenetic trees suggested that local outbreaks have become smaller in Japan; however, intermixing of viral types between these 2 areas was more evident in the more recent samples.
Subject(s)
Genotype , HIV Infections/epidemiology , HIV Infections/virology , HIV-1/classification , HIV-1/genetics , Adolescent , Adult , Aged , Blood/virology , Blood Donors , Cluster Analysis , Female , HIV-1/isolation & purification , Humans , Japan/epidemiology , Male , Middle Aged , Molecular Epidemiology , Phylogeny , RNA, Viral/genetics , RNA, Viral/isolation & purification , Sequence Analysis, DNA , Young Adult , env Gene Products, Human Immunodeficiency Virus/genetics , pol Gene Products, Human Immunodeficiency Virus/geneticsABSTRACT
Background: Most of the Japanese population is seropositive for anti-Japanese encephalitis virus (JEV) antibodies because of previous JEV vaccination or natural infection. Because the virological characteristics of JEV are similar to those of West Nile virus (WNV) and dengue virus (DENV), we hypothesized that anti-JEV antibodies can cross-react with WNV and DENV antigens, leading to protection against infection by these viruses. Methods: Using isolated intravenous immunoglobulin (IVIG) from plasma collected in Japan, neutralizing activities against WNV and DENV and antibody-dependent enhancement (ADE) of these viral infections were evaluated using an in vitro assay to determine the potency of immunity against these viruses. Results: The prepared IVIG showed considerable neutralizing activity of 2.57 log10 reduction factor against WNV infection but showed little effect against DENV infection. A strong correlation was observed between the neutralizing activity of individual plasma samples against JEV and WNV (ρ=0.768). Moreover, IVIG showed no significant ADE of WNV infection. Conclusions: Based on these results, we presume that the Japanese population is generally protected from WNV infection. Furthermore, IVIG prepared from plasma donations from Japanese individuals is expected to be an effective therapeutic agent based on its neutralizing activity against JEV and WNV.
Subject(s)
Antibodies, Neutralizing/blood , Dengue Virus/immunology , Dengue/prevention & control , Encephalitis Virus, Japanese/physiology , Encephalitis, Japanese/immunology , Plasma/immunology , West Nile Fever/prevention & control , West Nile virus/immunology , Animals , Antibodies, Viral/blood , Culicidae/immunology , Humans , Immunoglobulins , Japan , Neutralization TestsABSTRACT
BACKGROUND: Dengue fever is caused by four related RNA viruses of the genus Flavivirus, dengue virus (DENV)-1, -2, -3, and -4, which are transmitted to humans by mosquitoes. Although DENV is not endemic in Japan, an autochthonous dengue outbreak occurred in 2014. Several transfusion-transmitted cases have also been reported after the use of blood and plasma products in DENV-endemic countries. The aim of this study was to develop a novel multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay for DENV blood screening. STUDY DESIGN AND METHODS: Large-scale oligonucleotide screening was performed to obtain DENV-specific primers and probes using a variety of DENV clinical isolates. A multiplex RT-PCR assay was then developed using the identified oligonucleotides and the ability of this assay to detect DENV RNA was evaluated. RESULTS: A number of oligonucleotides suitable for DENV RNA detection were identified and a novel DENV serotype-specific multiplex RT-PCR assay was successfully established. Comparative analysis revealed that the multiplex assay could detect levels of viral contamination as low as 100 viral copies/mL. CONCLUSION: This established serotype-specific multiplex RT-PCR assay provides a simple, sensitive, and quantitative detection method for DENV, which could be applied in the screening of blood samples to prevent transfusion-transmitted DENV infection.
Subject(s)
Dengue Virus/genetics , Dengue/diagnosis , Polymerase Chain Reaction/methods , Serogroup , Transfusion Reaction , Blood Safety , Dengue/prevention & control , Dengue/transmission , Humans , Multiplex Polymerase Chain Reaction , RNA, Viral/analysis , RNA, Viral/blood , Real-Time Polymerase Chain ReactionABSTRACT
BACKGROUND: Bacterial contamination of platelet concentrates (PCs) remains a serious problem in transfusion. We have been conducting sterility tests on all PCs rejected by blood centers or hospitals due to abnormal appearances. We recently experienced a case in which discrepant results were obtained between the methods used to identify a bacterial species isolated from a PC, requiring further analyses. STUDY DESIGN AND METHODS: Bacteria were isolated from a PC using the BacT/ALERT system and plate culture. The species was identified using biochemical tests and molecular analysis. Phylogenetic trees were constructed using sequences of the 16S ribosomal RNA (rRNA) and superoxide dismutase (sodA) genes from the bacterial isolate and related species. In addition, the isolate was cultured at temperatures of 10°C and below to determine its growth activity at low temperatures. RESULTS: Biochemical tests determined that the isolate was Streptococcus alactolyticus, whereas molecular analysis determined that it was Lactococcus garvieae. These two species belonged to different clusters on the phylogenetic tree. Similar to L. garvieae, the isolate could grow at 10°C. CONCLUSIONS: We conclude that the isolate was L. garvieae according to molecular identification and its growth characteristic at 10°C. Molecular analysis enabled the identification of this species, which was difficult to classify by biochemical tests. Blood facilities need to be prepared with multiple techniques, including genetic analysis techniques, for identifying contaminating bacterial species. L. garvieae can grow at 10°C and can contaminate both red blood cell concentrates and PCs; thus, this species should be listed as a cryophilic bacterium that could threaten blood safety.
Subject(s)
Blood Platelets/microbiology , Lactococcus/genetics , Lactococcus/isolation & purification , Blood Safety , Genes, Bacterial , Humans , Japan , Molecular Typing/methods , Phylogeny , Platelet Transfusion , RNA, Ribosomal, 16S/genetics , Superoxide Dismutase/genetics , TemperatureABSTRACT
Quantitative PCR (qPCR) analysis of human T-cell leukemia virus type 1 (HTLV-1) was used to assess the amount of HTLV-1 provirus DNA integrated into the genomic DNA of host blood cells. Accumulating evidence indicates that a high proviral load is one of the risk factors for the development of adult T-cell leukemia/lymphoma and HTLV-1-associated myelopathy/tropical spastic paraparesis. However, interlaboratory variability in qPCR results makes it difficult to assess the differences in reported proviral loads between laboratories. To remedy this situation, we attempted to minimize discrepancies between laboratories through standardization of HTLV-1 qPCR in a collaborative study. TL-Om1 cells that harbor the HTLV-1 provirus were serially diluted with peripheral blood mononuclear cells to prepare a candidate standard. By statistically evaluating the proviral loads of the standard and those determined using in-house qPCR methods at each laboratory, we determined the relative ratios of the measured values in the laboratories to the theoretical values of the TL-Om1 standard. The relative ratios of the laboratories ranged from 0.84 to 4.45. Next, we corrected the proviral loads of the clinical samples from HTLV-1 carriers using the relative ratio. As expected, the overall differences between the laboratories were reduced by half, from 7.4-fold to 3.8-fold on average, after applying the correction. HTLV-1 qPCR can be standardized using TL-Om1 cells as a standard and by determining the relative ratio of the measured to the theoretical standard values in each laboratory.
Subject(s)
DNA, Viral/analysis , Human T-lymphotropic virus 1/genetics , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/standards , Viral Load/genetics , Cell Line, Tumor , DNA, Viral/genetics , HTLV-I Infections/genetics , HTLV-I Infections/virology , Humans , Japan , Jurkat Cells , Leukemia, T-Cell/genetics , Leukemia, T-Cell/virology , Leukocytes, Mononuclear/virology , Proviruses/genetics , Virus Integration/geneticsABSTRACT
BACKGROUND: It has been demonstrated that the hepatitis E virus (HEV) can be transmitted via blood transfusion, and the risk of HEV transmission via transfusion has become a major global concern. An HEV culture system for blood-derived HEV has been sought to obtain valuable knowledge of the virus and the risk of HEV infection through blood products. STUDY DESIGN AND METHODS: We endeavored to establish an HEV culture system using RNA-positive blood specimens for Genotypes (G) 3 and 4 and applied this system to evaluate tissue culture infectious dose (TCID). We applied this method to investigate the potential of the Mirasol pathogen reduction technology (PRT) system (Terumo BCT) to inactivate live HEV in contaminated platelet samples (PLTs). PLTs were spiked with cultured HEV G3 or G4 and then treated with the Mirasol PRT system. PLTs were examined before and after the treatment for HEV load using TCID titration. RESULTS: We successfully established two strains for HEV production: the JRC-HE3 strain for G3 and the UA1 strain for G4. The Mirasol PRT system expressed more than 3 log inactivation for JRC-HE3 and more than 2 log inactivation for UA1. CONCLUSION: The Mirasol PRT system inactivated greater than 2 to 3 logs of live HEV in PLTs and can potentially be used to lower the possibility of blood-borne HEV transmission. The G3 and G4 HEV inocula identified in this study and the hepatoma cell culture system provide a new means to assess HEV infectious titer and to evaluate other pathogen reduction strategies.
Subject(s)
Genotype , Hepatitis E virus/growth & development , Virus Inactivation , Cell Line, Tumor , Hepatitis E virus/isolation & purification , HumansABSTRACT
Pemphigoid gestationis (PG) is a rare, perinatal, autoimmune, and blistering dermatosis. Only few cases of PG involving hydatidiform moles have been reported. Complete hydatidiform moles are usually evacuated by dilatation and curettage. We report a patient with a massive complete hydatidiform mole that underwent spontaneous expulsion; she subsequently developed PG. A 19-year-old unmarried nulligravid woman was referred to our hospital following excessive vaginal bleeding after an uncertain amenorrheal period. The patient presented with preshock vital signs, severe anemia, and a positive urine pregnancy test. Imaging examinations revealed a massive intrauterine mass (19 × 15 × 10 cm), suggesting a complete hydatidiform mole. She was hospitalized and treated with blood transfusion. Sixteen hours after hospitalization, the massive molar mass underwent spontaneous expulsion and bleeding ceased. Three days after the expulsion, she developed pruritic skin lesions including papules, erythemas, and bullae, which spread over her entire body. Skin biopsy revealed PG and subepidermal blister formation and linear complement C3 deposition along the basement membrane zone, and the serum anti-BP180 antibody level was found to be high on measurement. She was effectively treated with 50 mg/day of oral prednisolone. Her skin lesions disappeared, leaving pigmentation.
ABSTRACT
The xenotropic murine leukemia virus-related virus (XMRV) was first described as a novel human gammaretrovirus in prostate tumor tissues and was reported to be found in blood, suggesting the possibility of XMRV transmission via blood transfusion. The gag and env regions of the XMRV proviral DNA that were detected 1,030 blood samples collected from the greater Tokyo area were examined by real-time PCR analysis. However, XMRV infection was not found in the samples; this suggested that the risk of XMRV transmission via transfusion is very low in Japan.
Subject(s)
DNA, Viral/blood , Proviruses/genetics , Xenotropic murine leukemia virus-related virus/genetics , Adolescent , Adult , Aged , Animals , Blood Donors , Female , Genes, env , Genes, gag , Genome, Human , Humans , Male , Mice , Middle Aged , Virus Integration , Young AdultABSTRACT
BACKGROUND: In most cases of anaphylactic transfusion reaction, the mechanisms underlying its development are unclear. We found a donor whose transfused blood components were implicated in two cases of anaphylactic transfusion reaction, and we found that the donor plasma showed mast cell degranulation activity. STUDY DESIGN AND METHODS: The donor plasma was examined to identify the mast cell-activating factors in it. Cultured mast cells prepared from cord blood were used for in vitro degranulation assay. Serum prepared from the donor plasma was fractionated by three-step chromatography using mast cell degranulation activity as a marker. The fractions selected from the third step of chromatography were analyzed by mass spectrometry after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The characteristics of the identified proteins and other plasma samples that had been donated by the donor over several years were examined. RESULTS: Two plasma proteins of high molecular weight were detected in the anion-exchange fractions and identified as human immunoglobulin (Ig)Es of 500 kDa and higher. The mast cell degranulation activity of the IgEs decreased in the presence of monomeric human IgE as well as an anti-human IgE antibody. Mast cell degranulation activity was detected in the donor plasma since January 4, 2002, when the first case was reported. CONCLUSION: We identified high-molecular-weight IgEs as the mast cell-activating factors in the donor plasma. Results of analysis suggest that these IgEs were dimeric and trimeric and that they directly activated the transfusion recipient's mast cells by triggering the crosslinking of Fcε receptor I, thereby inducing an anaphylactic transfusion reaction.
Subject(s)
Anaphylaxis/etiology , Immunoglobulin E/blood , Mast Cells/physiology , Protein Multimerization , Transfusion Reaction , Adult , Aged , Blood Donors , Cell Degranulation , Female , Humans , Immunoglobulin E/immunology , Male , Middle Aged , Molecular WeightABSTRACT
BACKGROUND: In the spring of 2009, the novel swine-origin influenza A (pandemic [H1N1] 2009) virus emerged and spread globally. Although no established cases of transfusion-transmitted influenza have been reported, the widespread outbreak of pandemic (H1N1) 2009 caused serious concern regarding the safety of blood products. The Japanese Red Cross Blood Centers have intercepted blood products with accompanying postdonation information indicating possible pandemic (H1N1) 2009 infection. To study the risk of transmission of pandemic (H1N1) 2009 by blood transfusion, we searched for the viral genome in such products using nucleic acid amplification technology. STUDY DESIGN AND METHODS: Between June and December 2009, blood components were collected from 579 blood donors who were diagnosed as or strongly suspected of having pandemic (H1N1) 2009 within 7 days after donation. Viral RNA was extracted from plasma and red blood cell (RBC) products, and RNA samples were subjected to real-time reverse transcription-polymerase chain reaction of the hemagglutinin and matrix genes of the pandemic (H1N1) 2009 virus. RESULTS: A total of 565 plasma and 413 RBC products from the 579 blood donors were available. No viral RNA of the pandemic (H1N1) 2009 was detected in any of the blood samples from the 579 blood donors. CONCLUSION: No viremia of pandemic (H1N1) 2009 was demonstrated in any of the 579 blood donors who had most likely donated blood during the incubation period. It is considered that the risk of transmitting pandemic (H1N1) 2009 by blood transfusion is extremely low.