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Biomedical subjects

P Goubau

Publications and source records attributed to P Goubau.

At least 37 records · Page 2Linked to original sources

Epstein-Barr virus infection in sixty pediatric liver graft recipients: diagnosis of primary infection and virologic follow-up.

BACKGROUND: Epstein-Barr virus (EBV) is an important cause of infection after pediatric liver transplantation. Earlier detection of EBV could result in shortening the delay in diagnosis and allow better management of a pediatric high risk population. OBJECTIVES: To determine the timing of EBV primary infection after graft and to compare the performances of different assays for an early detection of the virus. METHODS: Sixty pediatric liver graft recipients were followed. Kinetics of appearance of different EBV serologic parameters (anti-EBV-IgG, -IgM and -IgA, and anti-EBV nuclear antigen-IgG) and of the viral DNA in peripheral blood lymphocytes by PCR were compared. RESULTS: Thirty-six patients had a primary EBV infection. The first PCR and IgM positive result appeared after a mean delay of 56 and 61 days, respectively, and preceded the IgG response (mean delay, 143 days). Most of the studied patients (13 of 16) developed anti-EBV-IgA and only 3 developed anti-EBNA-IgG during the follow-up period. CONCLUSIONS: EBV primary infection occurred in most cases during the first 2 months after graft. The IgG response was delayed. The best performance was obtained by PCR. However, the IgM test compared well with the PCR and could be a more widely accessible measure to follow regularly.

Adolescent↗

Increased risk of cytomegalovirus transmission in utero during late gestation.

OBJECTIVE: To determine whether the rate of human cytomegalovirus transmission in utero is related to the gestational age at the time of maternal infection. METHODS: One hundred twenty-three pregnant women followed in our units between 1988 and 1998 were studied retrospectively. Each had developed a primary infection with cytomegalovirus evidenced by a seroconversion, confirmed by specific enzyme immunoassays. Infants were diagnosed by urine culture. RESULTS: Regardless of gestational age at the time of maternal cytomegalovirus seroconversion, the mean rate of intrauterine transmission was 57.5%. There was a statistically significant difference between early seroconversion (during the first trimester) and late seroconversion (during the third trimester) (36.0% versus 77.6%; P < .001). The risk of transmission calculated for seroconversion during the second trimester was intermediate (44.9%). CONCLUSION: A statistically significant difference in the rate of intrauterine cytomegalovirus transmission was observed according to the duration of pregnancy at which primary infection occurred. The rate of transmission increased with gestational age.

Adult↗

Two new human T-lymphotropic virus type I phylogenetic subtypes in seroindeterminates, a Mbuti pygmy and a Gabonese, have closest relatives among African STLV-I strains.

Six new HTLV-I strains from seroindeterminate individuals were analyzed: four from Gabon, one from a Mbuti Efe pygmy in Congo (formerly Zaire), and one from a Congolese patient residing in Belgium. The LTR and env regions were sequenced and phylogenetic analyses were performed to characterize the new strains. Nucleotide divergence and phylogeny results showed that four of the new strains belong to the HTLV-Ib Central African subtype. The other two strains, one from the Efe pygmy and one from Gabon, lie on distinct branches of the LTR and env trees with respect to the four major HTLV-I subtypes. Despite the low bootstrap values, likelihood mapping analyses proved that these strains can be considered two new HTLV-I molecular subtypes, putatively named HTLV-Ie and HTLV-If. A relation exists in the phylogenetic trees and in the likelihood maps between the new subtypes and African STLV-I strains from Papio spp. and Cercopithecus spp., suggesting one or more interspecies transmission events in the past. This study demonstrates that the phylogenetic subtyping of HTLV-I in the African continent is far from being completed and that samples presenting an indeterminate serology can potentially belong to new subtypes in humans. In addition, present day serological tests do not reliably type strains within the HTLV-Ib Central African subtype.

Base Sequence↗

The simian T-lymphotropic virus STLV-PP1664 from Pan paniscus is distinctly related to HTLV-2 but differs in genomic organization.

We have isolated a highly divergent simian T-lymphotropic virus, STLV-PP1664, from a wild-caught bonobo (Pan paniscus). Previous phylogenetic analysis suggested that this virus represents an additional type of STLV but this has now become a matter of discussion. We have now obtained and analyzed the entire genome of STLV-PP1664. All major genes and their corresponding viral messengers were identified. Sequence comparison and phylogenetic analysis indicated that this virus, together with the closely related panp isolate, belongs to an early lineage within the PTLV-2 clade, differing from HTLV-2 by about 25%. In contrast to the HTLV-1 and HTLV-2 LTR, only two 21-bp repeats instead of three were found in the STLV-PP1664 LTR. Additional messengers, resulting from alternative splicing, potentially encode five different accessory proteins from open reading frames in the pX region: prorfI, porfII, ptorfV', and two isoforms of Rex. The amino acid sequences of these proteins are only distinctly related to the accessory proteins from HTLV-2. These data suggest a different genomic organization of the STLV-PP1664 pX region than that of HTLV-2. We conclude that STLV-PP1664, although related to HTLV-2, has some distinct features in the LTR and the pX regions, the impact of which needs further investigation. Although arguments pro and contra a distinct classification are nearly equally balanced, we propose to classify this virus as an STLV-2, designated STLV-2PP1664.

Amino Acid Sequence↗

Evolutionary rate and genetic heterogeneity of human T-cell lymphotropic virus type II (HTLV-II) using isolates from European injecting drug users.

Seven new Italian and two new British HTLV-II isolates were obtained from injecting drug users and the entire long terminal repeat (LTR) region was sequenced. Restriction analysis showed that all the Italian isolates are of the IIb subtype, whereas the British isolates are of the IIa subtype. To understand whether the further differentiation of each two principal HTLV-II subtypes in several subgroups could be statistically supported by phylogenetic analysis, the neighbor-joining, parsimony, and maximum likelihood methods were used. The separation between IIa and IIb is very well supported by all three methods. At least two phylogenetic subgroups exist within the HTLV-IIa and at least three within the HTLV-IIb subtype. In the present analysis, no statistical support was obtained for additional phylogroups. Two particular subgroups seem interesting because they include all European and North American injecting drug user strains within the IIa and IIb subtypes, respectively. These data confirm that European HTLV-II infection among drug users is probably derived from North America. They also suggest that though a certain differentiation by restriction analysis in different subgroups is possible, carefully interpreted phylogenetic analyses remain necessary. Using the likelihood ratio test, a molecular clock for the drug user strains was calibrated. A fixation rate between 1.08 x 10(-4) and 2.7 x 10(-5) nucleotide substitutions per site per year was calculated for the IIa and IIb injecting drug user strains. This is the lowest fixation rate so far reported for RNA viruses, including for HIV, which typically range between 10(-2) and 10(-4).

Biological Evolution↗

Avidity of IgG antibodies distinguishes primary from non-primary cytomegalovirus infection in pregnant women.

BACKGROUND: Human cytomegalovirus (HCMV) is the most common cause of viral intrauterine infection. Fetal damage is mostly linked to maternal primary infection. It is therefore important to differentiate primary from non-primary infection in pregnant females. IgM tests often used for this purpose are not reliable enough. OBJECTIVE: To evaluate an HCMV-IgG urea-elution assay for its ability to distinguish primary from non-primary infection. In this assay, soaking the antigen-antibody complex with an urea containing solution frees antibodies with low avidity but has no influence on those with high avidity. An avidity index (AI) was calculated: AI = (OD with urea/OD without urea) x 100. STUDY DESIGN: HCMV-IgG avidity was measured on a single serum of 79 patients with past infection (pregnant women, graft recipients and blood donors) and of 63 patients (78 sera) with documented seroconversion (pregnant women and graft recipients). Sixty-one pregnant women positive or equivocal for HCMV-IgM but without a documented seroconversion were included in this study. RESULTS: Most (72/79) of the patients with past infection had an AI > 65% and all but one had an AI > 50%. In pregnant women, in the case of a primary infection within the past 3 months, AI are usually (51/53) < 50% and never > 65%. Among the IgM positive pregnant women who lack a seroconversion history, 38 had AI > 65% suggestive of an infection that had occurred at least 3 months earlier, 11 had an AI in a grey area between 50 and 65% and 12 had an AI < 50%, suggestive of a recent primary infection. CONCLUSIONS: In pregnant women, measurement of the IgG avidity may help to date a HCMV infection, an AI > 65% highly suggests a past infection while an AI < 50% corresponds to a recent primary infection.

Antibody Affinity↗

GB virus C: a novel pathogen or a curiosity for virologists?

The recently identified GB virus C is a flavivirus related to the hepatitis C virus. It appears quite prevalent in the general population and especially in individuals at risk of parenteral transmission. Many questions remain, most importantly the pathogenic role of the virus. It is unlikely that this virus has a major role in hepatic disease.

Flaviviridae↗

African origin of human T-lymphotropic virus type 2 (HTLV-2) supported by a potential new HTLV-2d subtype in Congolese Bambuti Efe Pygmies.

We identified a potential new subtype within human T-cell lymphotropic virus type 2 (HTLV-2), HTLV-2d, present in members of an isolated Efe Bambuti Pygmy tribe. Two of 23 Efe Pygmies were HTLV-2 seropositive, with HTLV-2 Western blot and enzyme-linked immunosorbent assay reactivities. From one of them the entire genome of the HTLV-2 strain Efe2 could be amplified and sequenced. In all gene regions analyzed, this strain was the most divergent HTLV-2 strain, differing by 2.4% (tax/rex) to 10.7% (long terminal repeat) from both subtypes HTLV-2a and HTLV-2b, yet major functional elements are conserved. The similarity between the HTLV-2 Efe2 Gag and Env proteins and the corresponding HTLV-2a and -2b proteins is consistent with the observed serological reactivity. In the proximal pX region, one of the two alternative splice acceptor sites is abolished in HTLV-2 Efe2. Another interesting feature of this potential new subtype is that it has a Tax protein of 344 amino acids (aa), which is intermediate in length between the HTLV-2a Tax protein (331 aa) and the HTLV-2b and -2c Tax proteins (356 aa) and similar to the simian T-cell lymphotropic virus type 2 (STLV-2) PP1664 Tax protein. Together these two findings suggest a different phenotype for the HTLV-2 Efe2 strain. Phylogenetic analyses confirmed that the Pygmy Efe2 strain potentially belonged to a new and quite divergent subtype, HTLV-2d. When the STLV-2 bonobo viruses PP1664 and PanP were used as an outgroup, it was clear that the Pygmy HTLV-2 Efe2 strain had the longest independent evolution and that HTLV-2 evolution is consistent with an African origin.

Amino Acid Sequence↗

Molecular analysis of GB virus C isolates in Belgian hemodialysis patients.

GB virus C (GBV-C) has been detected in Belgian hemodialysis patients. To study their genomic diversity and phylogenetic relationship, a 592 nucleotide fragment extending from the 5' non-coding region to part of the E1 gene of the GBV-C genome was amplified and sequenced from 12 Belgian hemodialysis patients in two different centers. Together with strains from different geographical origins, these sequences were analyzed phylogenetically using three different methods. A consistent tree topology was obtained with all methods. Three GBV-C genotypes were observed with two subtypes in type 2 and a questionable subtyping in type 1. Except for one isolate falling into type 1 cluster which mainly consists of African strains, all the other Belgian strains clustered within the type 2a branch. Two GBV-C isolates in two patients from the same hemodialysis center clustered together closely, suggesting a nosocomial transmission. In view of their long branch length, it seems unlikely that the other Belgian strains evolved recently from a common ancestor. Our results indicate that the major type circulating among Belgian hemodialysis patients seems to be 2a, which is usual for Europe and North America, but that the African type 1 also exists to a minor extent. Although patient to patient transmission of GBV-C in Belgian hemodialysis centers did occur, it may not account for the majority of infections.

Base Sequence↗

Tympano-ossicular allografts and HIV transmission.

OBJECTIVE: To evaluate the potential risk of human immunodeficiency virus (HIV) transmission by tympano-ossicular allografts by studying the efficacy of standard preservation techniques to eliminate the presence of proviral HIV-1 DNA fragments in contaminated ossicles. STUDY DESIGN: Randomized single-blind prospective study on the ossicles of HIV-1 patients. MATERIAL: Ossicles of five patients who had died of acquired immune deficiency syndrome (AIDS) (HIV-1 infection) were taken within 6 hours postmortem and allocated randomly to a treatment and nontreatment group. Liver and skin biopsies were taken as positive control specimens. PROCESSING: The treatment group was processed with standard techniques (formaldehyde) for tympano-ossicular allograft preservation and the nontreatment group was only washed, dried, and stored in sterile tubes at -700 degrees without further processing. MAIN OUTCOME MEASURE: Proviral HIV-1 DNA was detected using polymerase chain reaction amplification techniques. RESULTS: No proviral HIV-1 DNA was detected in any of the treated ossicles, whereas three of five sets of untreated ossicles were positive. The positive control specimens of all treated and nontreated sets were positive for proviral HIV-1 DNA. CONCLUSIONS: These results suggest that the preservation technique for tympano-ossicular allografts is safe with regard to HIV-1 transmission.

DNA, Viral↗

Toxic shock syndrome without rash in a young child: link with syndrome of hemorrhagic shock and encephalopathy?

A diffuse macular erythroderma and subsequent desquamation after 1 to 2 weeks are two of the five major diagnostic criteria of toxic shock syndrome (TSS). We present the case of a 15-month-old girl with TSS, but without erythroderma or desquamation. She was admitted with high fever, shock, and multiorgan involvement. Minimal or no cutaneous signs were present. Initially the diagnosis of the syndrome of hemorrhagic shock and encephalopathy was made. After 7 days, a TSS toxin 1-producing strain of Staphylococcus aureus was cultured from an inguinal lymph node, where inflammation had already been noticed on admission. Moreover, the girl had no antibodies against this toxin. The serum cytokine profile during the acute phase of her illness showed high levels of tumor necrosis factor-alpha, interleukin-6 and interferon-gamma, as is seen during activation of the immune system by TSS toxin 1. Other possible causes for the patient's illness were excluded. We conclude that the patient had TSS without rash. Without the evidence implicating a TSS toxin 1-producing strain of S. aureus as the cause of her disease, a diagnosis of syndrome of hemorrhagic shock and encephalopathy would have been made. It is possible that some cases of syndrome of hemorrhagic shock and encephalopathy represent a variant of TSS in small children.

Bacterial Toxins↗

Comparative evaluation of three ELISA techniques and an indirect immunofluorescence assay for the serological diagnosis of Epstein-Barr virus infection.

BACKGROUND: The reference method for detecting specific Epstein-Barr virus (EBV) antibodies is indirect immunofluorescence (IF) with EBV-infected cells. The availability of protein purified from infected cells and more recently of recombinant polypeptides designed to contain immunodominant epitopes, has enabled the development of commercial enzyme-linked immunosorbent assays (ELISA) for the specific serodiagnosis of EBV infection. OBJECTIVE: Evaluation of ELISA-based EBV serodiagnosis in comparison with indirect immunofluorescence. STUDY DESIGN: We have first compared three commercial ELISA test systems with our in house indirect immunofluorescence assay for classifying correctly a set of serum samples into clinical categories (acute infection, past infection, interfering non-EBV infection, persistent infection). Additionally a prospective analysis with the best performing ELISA test (Enzygnost) was then carried out by running the ELISA test in parallel with the indirect immunofluorescence assay on 324 consecutive clinical samples sent to our laboratory for EBV serodiagnosis. RESULTS: For the serodiagnosis of past EBV infection and acute EBV infection all three commercial ELISAs performed well in comparison with indirect immunofluorescence. When testing samples positive for cytomegalovirus (CMV), Toxoplasma or herpes simplex IgM, interference in the IgM tests was observed with the three ELISAs. In some instances we could demonstrate that the positive IgM results were due to EBV reactivation. The observed discrepancies between ELISA and IF for the serodiagnosis of chronic EBV infection or EBV reactivation, point to the difficulty for the serodiagnosis of persistent EBV infection on single serum samples. According to our prospective study the EBV IgG determination was accurate. A positive IgM result was not always indicative of an acute infection. Positive IgM results due to EBV reactivation were observed. A positive EBV nuclear antigen (EBNA) IgG result in those samples precluded acute infection. CONCLUSIONS: 90-95% of samples could be classified correctly into clinical categories by a two parameter ELISA system detecting IgG and IgM against a standardized mixture of EBV antigens, allowing standardization and automation of EBV-specific serology. The absence of EBNA IgG was useful as a second line confirmatory assay for acute EBV infection.

Antibodies, Viral↗

Confirmatory strategy of hepatitis C serology based on two screening assays in a diagnostic setting.

One hundred ninety seven successive sera, positive for anti-hepatitis C virus antibody with a third generation screening enzyme immunoassay (MEIA on IMX, Abbott), were tested with alternative third generation screening assays from two different manufacturers (Sanofi and Ortho), with an immunoblot assay (RIBA 3.0, Chiron), and by reverse transcriptase polymerase chain reaction (RT-PCR). Samples positive by RIBA 3.0 or by RT-PCR were considered as true positives. The positive predictive value of a combination of strong positive results in two screening assays was more than 98%. This combination of results has the same predictive value for detectable viraemia as a positive RIBA 3.0 (86.5%). Using a policy of two successive enzyme immunoassays in this clinical diagnostic setting diminishes the need for supplemental assays by more than 85%.

Hepatitis C↗

Hepatitis G virus infection in haemodialysed patients: epidemiology and clinical relevance.

BACKGROUND: The prevalence, incidence, risk factors, and clinical impact of infection by the recently discovered hepatitis G virus (HGV) in haemodialysed (HD) patients, are poorly defined. METHODS: All 119 HD patients from two Belgian units selected for their different hepatitis C virus (HCV) prevalences (A: 19.2%, B: 3.4%) were tested for the presence of HGV-RNA, using the reverse transcriptase polymerase chain reaction (RT-PCR) and primers from the 5'-NC and NS 5a genome regions. The results of anti-HCV antibodies and alamine aminotransferase levels (ALT) at the time of RT-PCR, number of transfusions from the onset of HD, and time on HD were retrieved from the medical charts. Forty patients were retested by RT-PCR 3-64 months later. RESULTS: HGV-RNA was detected with both sets of primers in 11/78 patients (14.1%) from centre A and 8/41 patients (19.5%) from centre B, for an average prevalence of 16%. One patient was indeterminate (positive with one set of primers). The presence of HGV-RNA correlated neither with time on HD (P = 0.18), nor with the number of transfusions on HD (P = 0.14). It was associated with the presence of anti-HCV antibodies in centre A (P < 0.01) but not B (P > 0.5). Twenty-seven initially negative (-) patients (A: n = 18; B: n = 9) were retested: two became positive (+) both in the absence of transfusions for years, giving a yearly incidence of 1.7%. The 13 initially HGV-RNA (+) patients remained so over time (33 patient-years). The presence of HGV-RNA alone does not increase significantly the ALT level, in contrast to the strong influence of HCV. CONCLUSION: The prevalence and yearly incidence of HGV infection are 16% and 1.7%, respectively, in our HD patients. Neither the number of transfusions on HD nor the time on HD are significant risk factors. Although mixed HCV/HGV infections indicate common risks, the prevalence of HCV in a particular setting does not predict prevalence of HGV. As new infections are detected in the absence of blood transfusions, HGV may be another marker of nosocomial viral transmission. Once acquired, the infection persists for many years in HD patients.

Adult↗

Complete nucleotide sequence of the new simian T-lymphotropic virus, STLV-PH969 from a Hamadryas baboon, and unusual features of its long terminal repeat.

A third type of primate T-lymphotropic virus, PTLV-L, with STLV-PH969 as a prototype, has recently been isolated from an African baboon (Papio hamadryas). Classification of this virus has been based on partial sequence analysis of cDNA from a virus-producing cell line, PH969. We obtained the complete nucleotide sequence of this virus with a proviral genome of 8,916 bp. All major genes, homologous in all human T-cell lymphotropic virus (HTLV)-related viruses, and their corresponding mRNAs, including appropriate splicing, were identified. One additional nonhomologous open reading frame in the proximal pX region is accessible for translation through alternative splicing. Sequence comparison shows that STLV-PH969 is equidistantly related to HTLV type 1 (HTLV-1) and HTLV-2. In all coding regions, the similarity tends to be the lowest between STLV-PH969 and HTLV-1. However, in the long terminal repeat (LTR) region, the lowest similarity was found between STLV-PH969 and HTLV-2. The U3-R and R-U5 boundaries of the STLV-PH969 LTR were experimentally determined at nucleotides 268 and 524, respectively. This 695-bp LTR is 60 and 73 bp shorter than the LTRs of HTLV-1 and HTLV-2, respectively, but its general organization is similar to the one found in the HTLV-bovine leukemia virus genus. In the long region between the polyadenylation signal and the poly(A) site, sequence similarity with the HTLV-1 Rex-responsive element (RexRE) core and secondary structure prediction suggest the presence of a RexRE. The presence of three 21-bp repeats is conserved within the U3 region of HTLV-1, HTLV-2, and BLV. Only two direct repeats with similarity to these Tax-responsive elements were found in the STLV-PH969 LTR, which might suggest differences in the Tax-mediated transactivation of this virus. We conclude that STLV-PH969 has all the genes and genomic regions to suggest a replication cycle comparable to that of HTLV-1 and HTLV-2.

Amino Acid Sequence↗

Use of a generic polymerase chain reaction assay detecting human T-lymphotropic virus (HTLV) types I, II and divergent simian strains in the evaluation of individuals with indeterminate HTLV serology.

In countries with a low prevalence of human T-lymphotropic virus (HTLV) infection, indeterminate HTLV serologies are a major problem in blood bank screening because of the uncertainties about infection in these cases. The recent discovery of two new types of simian T-lymphotropic viruses (STLV), which give an HTLV-indeterminate serology, raises the question whether indeterminate serologies in humans may be linked to new types of HTLV. Starting from a Tax sequence alignment of all available primate T-cell lymphotropic virus strains (PTLV), including the two new types STLV-PH969 and STLV-PP1664, we developed generic and type-specific nested polymerase chain reactions (PCRs). The generic PCR proved to be highly sensitive and cross-reactive for all four types of PTLV, while the discriminatory PCRs had a high sensitivity and a specificity of 100%. There was no cross-reactivity with human immunodeficiency virus (HIV), ensuring correct interpretation of results from coinfected patients. Among the 77 serologically indeterminate samples tested, 6 were found to be HTLV-1 PCR positive and 1 was HTLV-II PCR positive. Sequencing of one of the HTLV-I PCR positives excluded PCR contamination, and revealed a divergent type of HTLV-I. The majority of the seroindeterminate samples (91%) were however HTLV-PCR negative, and no new types of HTLV were found. This new assay can identify otherwise undetected HTLV-I or HTLV-II infections and is a useful tool of screening for new types of HTLV among seroindeterminate samples.

Animals↗

Quantification of HIV-1 RNA in plasma: comparable results with the NASBA HIV-1 RNA QT and the AMPLICOR HIV monitor test.

We investigated and compared the reproducibility, accuracy, detection limits, and dynamic ranges of two commercial kits for quantification of RNA viral load using a titrated virus stock (laboratory strain HIV-1 IIIB) and 107 plasma samples of 25 HIV-1-infected patients. The high reproducibility of both methods (SD = 0.2-0.3 log for both methods) allowed reliable detection of a 0.5 log change in RNA viral load. Both methods had a similar detection limit (at least 10(3) RNA copies/ml plasma) and a dynamic range that extended over a 5 log (AMPLICOR) or a 6 log (NASBA) range of HIV-1 input. For HIV-1 IIIB, the viral load was compatible with measurements of virus-associated p24 antigen. For 21 patients (91 samples), the RNA viral load was similar with both methods differing by no more than 0.5 log. For four patients, the difference in viral load between the two methods was > 0.5 log for all 16 samples. For three of these patients, this could be explained by mismatches with primers or probes in the gag sequence: there was no correlation to the viral subtype. The RNA viral load determination was highly sensitive compared with p24 antigen measurement (> 95% of patients had a detectable viral load vs. 40% who had a detectable p24 level), but in the p24-positive samples the correlation between the antigen level and the RNA viral load was of only borderline significance. We also found that the viral RNA in whole blood was stable for at least 48 h during transport at room temperature. These observations show that both the NASBA HIV-1 RNA QT test and the AMPLICOR HIV monitor test are reliable parameters of the viral load, with great promise for their use as potential surrogate markers.

Base Sequence↗