SARS-CoV-2 testing, positivity, and factors associated with COVID-19 among people with HIV across Europe in the multinational EuroSIDA cohort.

BACKGROUND: Although people with HIV might be at risk of severe outcomes from infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; coronavirus 2019 [COVID-19]), regional and temporal differences in SARS-CoV-2 testing in people with HIV across Europe have not been previously described. METHODS: We described the proportions of testing, positive test results, and hospitalizations due to COVID-19 between 1 January 2020 and 31 December 2021 in the EuroSIDA cohort and the factors associated with being tested for SARS-CoV-2 and with ever testing positive. RESULTS: Of 9012 participants, 2270 (25.2%, 95% confidence interval [CI] 24.3-26.1) had a SARS-CoV-2 polymerase chain reaction test during the study period (range: 38.3% in Northern to 14.6% in Central-Eastern Europe). People from Northern Europe, women, those aged <40 years, those with CD4 cell count <350 cells/mm3, and those with previous cardiovascular disease or malignancy were significantly more likely to have been tested, as were people with HIV in 2021 compared with those in 2020. Overall, 390 people with HIV (4.3%, 95% CI 3.9-4.8) tested positive (range: 2.6% in Northern to 7.1% in Southern Europe), and the odds of testing positive were higher in all regions than in Northern Europe and in 2021 than in 2020. In total, 64 people with HIV (0.7%, 95% CI 0.6-0.9) were hospitalized, of whom 12 died. Compared with 2020, the odds of positive testing decreased in all regions in 2021, and the associations with cardiovascular disease, malignancy, and use of tenofovir disoproxil fumarate disappeared in 2021. Among study participants, 58.9% received a COVID-19 vaccine (range: 72.0% in Southern to 14.8% in Eastern Europe). CONCLUSIONS: We observed large heterogeneity in SARS-CoV-2 testing and positivity and a low proportion of hospital admissions and deaths across the regions of Europe.

[A disseminated strongyloidiasis in a non immunocompromised patient]. / Une strongyloïdose disséminée chez un patient immunocompétent.

Strongyloidiasis is a parasitosis mainly located in tropical and subtropical regions. Its European incidence is increasing due to population migration. The symptomatology is variable, largely depending on the host immune status. The diagnosis may be challenging as a specific inquiry is needed. The disseminated status, called hyperinfection syndrome, is a life-threatening disease that might occur in immunosuppressed patients. The patient's chances for a successful outcome depend on a quick and specific management. The treatment is based on anthelminthic agents such as ivermectine or albendazole for which the dosage will vary according to the infection type.

La strongyloïdose ou anguillulose est une parasitose principalement retrouvée dans les régions tropicales ou subtropicales. Son incidence en Europe est en augmentation suite aux flux migratoires. Sa symptomatologie est très variable et dépend fortement du statut immunitaire de l'hôte. Le diagnostic peut être difficile et nécessite une recherche spécifique. La forme disséminée, appelée hyperinfection, est typiquement présente chez les patients immunodéprimés. Elle est mortelle, surtout en l'absence de prise en charge adéquate. Le traitement est basé sur les antihelminthiques tels que l'ivermectine et l'albendazole dont la posologie est à adapter en fonction du type d'infection.

Effect of vitamin E supplementation of sheep and goats fed diets supplemented with polyunsaturated fatty acids and low in Se.

Vitamin E (VitE) and selenium (Se) are an essential part of the antioxidative functions of metabolism. There are situations of low supply of both micronutrients. As VitE is involved in ruminal biohydrogenation of polyunsaturated fatty acids (PUFA) and their protection against oxidation in metabolism, diets supplemented with PUFA may challenge VitE to an extent making recommended supplies insufficient. Twelve goats and sheep each were fed a diet supplemented with PUFA and characterised by low Se and limited VitE contents during the last 2 months of gestation and the first 2 months of lactation. The basal diet consisted of hay and concentrate. Six goats and sheep received extra VitE, while the control groups received no extra VitE. Blood and milk samples were taken. In addition, liver, heart muscle and spleen samples were obtained from the offspring after slaughtering at an age of 8 weeks. No significant changes were observed in serum Se and VitE. A significant increase in serum VitE concentrations between 2 and 4 weeks postpartum (pp) was evident in the supplemented kids. In 4, 6 and 8 weeks pp, the serum concentrations of VitE in the supplemented kids were significantly higher compared to the unsupplemented group. In the kids, VitE was higher in liver of the supplemented groups. There were no significant differences in response to extra VitE between sheep and goat. The kids responded to serum VitE different from that of lambs, as a significant difference was observed between supplemented and unsupplemented animals in the goat kids, but not the lambs. In conclusion, goats and sheep have to be viewed differently and may not be considered alike relating to VitE/Se metabolism and requirements, especially in young animals.

Response to antiretroviral therapy among patients exposed to three classes of antiretrovirals: results from the EuroSIDA study.

There is an increasing proportion of HIV-positive patients exposed to all licensed classes of antiretrovirals, and the response to salvage regimens may be poor. Among over 8500 patients in EuroSIDA, the proportion of treated patients exposed to nucleosides, protease inhibitors (PIs) and non-nucleoside reverse transcriptase inhibitor (NNRTI) increased from 0% in 1996 to 47% in 2001. Four-hundred-and-thirteen patients, who had failed virologically two highly active antiretroviral therapy (HAART) regimens and experienced all three main drug classes, started a salvage regimen of at least three drugs, in which at least one new PI or NNRTI was included. Median viral load was 4.7 log copies/ml [Interquartile range (IQR) 4.2-5.2], CD4 lymphocyte count 150/mm3 (IQR 60-274/mm3) and follow-up 14 months. Of these patients, 283 (69%) subsequently experienced at least a 1 log decline in viral load and 202 (49%) achieved a viral load < 500 copies/ml. Conversely, the CD4 count halved from the baseline value in 88 (21%), and 45 (11%) experienced a new AIDS-defining disease. In multivariable analyses, a 1 log viral load reduction was related to baseline viral load [relative hazard (RH) 1.27 per 1 log higher; P = 0.008], a previous viral load of less than 500 copies/ml (RH 1.69; P = 0.002), more recent initiation of the regimen (RH 1.36 per year more recent; P = 0.02), number of new drugs in the regimen (RH 1.20 per drug; P = 0.02), time since start of antiretroviral therapy (RH 0.94 per extra year; P = 0.035) and time spent on HAART with viral load > 1000 copies/ml (RH 0.96 per extra month; P = 0.0001). Analysis of factors associated with CD4 count decline and new AIDS disease also indicated improved outcomes in more recent times and a tendency for a better response in those starting more new drugs, but no relationship with the total number of drugs. Outcomes in people starting salvage regimens appear to depend on the number of new drugs started but not on the total number of drugs being used.

Genotypic correlates of resistance to HIV-1 protease inhibitors on longitudinal data: the role of secondary mutations.

Direct sequencing of the pol gene was assessed retrospectively with protease inhibitor susceptibility in a longitudinal study. A total of 134 samples from 26 patients were analysed at regular intervals up to 2 years. Patients were included in virological failure despite indinavir, ritonavir or saquinavir based triple-drug therapy. Both the type and number of certain secondary protease mutations modulated the effect of primary mutations on phenotypic resistance. This was notably applicable to L101/V, and to lesser extents to A711V/T. However, combinations of primary mutations, including 154V could predict resistance to the drug used and nelfinavir in more than 80%. In contrast, in vitro cross-resistance to amprenavir was rarely encountered. In addition, there was a relationship between a higher number of key mutations and poorer virological and clinical outcomes, respectively, from 6 and 3 months on. The key mutations were the protease mutations independently conferring phenotypic resistance and/or the reverse transcriptase mutations predicting treatment outcome. This relationship was independent from drug history, viral load and CD4 cell count measurements. In summary, even on a small sample size, sequence-based genotyping seems to be a good prognostic marker when performed longitudinally. In the context of primary resistance mutations, including additional secondary mutations, it may be useful in the prediction of phenotypic and clinical resistance. This should be assessed to optimize treatment monitoring before emergence of broadly cross-resistant virus.

Immunogenicity and safety of a new liquid hexavalent combined vaccine compared with separate administration of reference licensed vaccines in infants.

OBJECTIVE: The immunogenicity and safety of a new liquid hexavalent vaccine (diphtheria-tetanus-acellular pertussis-inactivated polio vaccine-hepatitis B-polyribosyl ribitol phosphate conjugated to tetanus protein; Hexavac; Aventis Pasteur MSD, Lyon, France) are compared with those of reference vaccines [diphtheria-tetanus-acellular pertussis-inactivated polio vaccine reconstituting lyophilized purified Haemophilus influenzae polysaccharide conjugated to tetanus protein vaccine (Pentavac; Aventis Pasteur MSD) and hepatitis B vaccine (H-B-Vax II; Aventis Pasteur MSD)] injected separately at the same visit in a prospective multicenter, comparative, open label trial. METHODS: Infants were randomized to receive Hexavac (n = 423) or Pentavac and H-B-Vax II (n = 425) as a primary immunization series at 2, 4 and 6 months of age. Seroprotection and seroconversion rates against all antigens at 1 month after the primary series were compared between the two vaccine groups with 95% confidence intervals (CI0.95) and were considered clinically equivalent (not inferior) when the upper limit of the 95% confidence interval on the difference (reference, hexavalent) was below predefined differences. RESULTS: Hexavac met and surpassed the pre-defined criteria for clinical equivalence to Pentavac and H-B-Vax II given concomitantly. It elicited similar seroprotection and seroconversion rates against all antigens. Seroprotection and seroconversion rates obtained 1 month after the third dose of Hexavac were >90% for all antigens. The postimmunization antibody geometric mean titers (GMT) for hepatitis B and purified Haemophilus influenzae polysaccharide were about 2-fold higher in infants who received the reference vaccines than in infants who had received Hexavac. GMTs for poliovirus antibodies tended to be enhanced in infants vaccinated with Hexavac. GMTs for all other antigens were very similar among both groups. Hexavac was generally well-tolerated. At least one local reaction was reported in 20.3% of Hexavac injections compared with 15.8% at the Pentavac injections site and 3.8% at the H-B-Vax II injections site. These reactions were generally mild and transient. At least one systemic adverse event was reported in 45.7% of Hexavac injections compared with 42.2% of Pentavac and H-B-Vax II injections (mild fever, irritability and drowsiness were most frequently reported). The frequency of adverse events was not significantly different between groups. No vaccine-related serious adverse event occurred during the study. CONCLUSION: This liquid hexavalent vaccine was generally well-tolerated and provided immune responses adequate to be protective against six infectious diseases with a single injection, given at 2, 4 and 6 months of age.

Three-year effectiveness of highly active antiretroviral treatment in the Luxembourg HIV cohort.

UNLABELLED: Clinical trials have shown that highly active antiretroviral treatment (HAART) is able to reduce HIV plasma viral loads to undetectable in 70% to 90% of patients and to increase CD4 cell counts. HAART in community settings (i.e., nonclinical trial situations) is reported to be much less effective. STUDY DESIGN: Observational study. PURPOSE: The aim of our study was to evaluate the effectiveness of protease inhibitor (PI)-based HAART in the Luxembourg HIV cohort after 36 months of treatment in previously treated and untreated patients. The secondary aim was to identify surrogate markers associated with long-term virologic and immunologic outcomes. PATIENTS AND METHOD: Seventy-three PI-naive patients, who started on HAART, combining one PI and two nucleoside reverse transcriptase inhibitors (NRTIs),with a follow-up of 3 years, were evaluated with plasma viral load and CD4 cell counts every 3 months and were analyzed retrospectively. Patients who had been treated previously with NRTI (n = 48) were at a more advanced stage of disease. RESULTS: Overall, there was a mean decrease in viral load compared to baseline of -1.89 log RNA copies/mL (SD = 1.40) that persisted at month 36. Sixty-two percent (62%) of patients reached an undetectable viral load (i.e., below 500 copies/mL): 82% and 53% of NRTI-naive and NRTI-experienced patients, respectively (p =.013). CD4 cell counts increased progressively in both groups with a sustained effect (mean increase of 146 cells/mL +/- 241) at month 36. NRTI-naive patients had a mean increase of 257 cells/mL (SD = 305), in contrast to experienced patients who had an increase of 108 cells/mL (SD = 206) at 3 years. Proportions of patients with a CD4 count under 200 cells/mL fell after 3 years for NRTI-naive (from 66% to 43%) and for experienced patients (from 32% to 13%). Predictors of short duration of viral load response were in decreasing order of importance: clinical AIDS, the use of saquinavir hard gel formulation as initial PI, and the number of NRTIs previously used. Viral load response was the only significant predictor of CD4 changes. CONCLUSION: In a community setting, effectiveness of PI-based HAART at 3 years is still achieved for most patients. NRTI-experienced patients have a good long-term response rate even if it is lower than NRTI-naive patients. A poor treatment response is associated with a more advanced stage of disease before HAART is introduced.

Maternal and perinatal health in Mali, Togo and Nigeria.

OBJECTIVE: The health care system in many developing countries is less efficient compared with that in the industrialized world. The aim of the present study was to investigate the differences of the efficiency of the health care in obstetrics in Nigeria, Mali and Togo. STUDY DESIGN: The data were collected in African district hospitals from Lomé/Togo (n = 1002), Bamako/Mali (n = 1462) and Kaduna/Nigeria (n = 1055) with a routinely applied questionnaire analyzed at the University Giessen and compared with the data from the Department of Obstetrics and Gynaecology Giessen (Germany) (referral hospital) (n = 1313) and the total data pool of the perinatal survey 1993 in Hesse (n = 58430). RESULTS: The medical history indicates important differences: the incidence of young mothers below 18 is highest in Mali (9.8%) compared with Nigeria (2.7%) and Togo (2.6%) (Hesse 0.6%). In general there is an increased rate of previous pregnancies of more than five: 13-28% (Hesse 2.1%). The rate of prenatal visits is also reduced: more than ten visits have 2% in Togo, 10.6% in Mali and 15.5% in Nigeria (Hesse 72%). Ultrasound examinations are very rare (1-12%) and tocolysis is nearly unknown. As a result the incidence of dead infants in the medical history is high: Mali 28%, Togo 18%, Nigeria 10.8% (Hesse 1.7%), as well as the rate of low birth weight infants. This is also reflected in the perinatal mortality which ranged from 115/1000 in Mali, 77/1000 in Togo and 68/1000 in Nigeria (Hesse 5.3/1000, Ob/Gyn Giessen 16.4/1000). CONCLUSION: The improvement of perinatal and maternal health in the developing world can only be achieved if family planning, prenatal care, selection of high risk pregnancies goes in parallel with a sound organization implemented and supported by the government.

[Rheumatoid purpura and intravenous immunoglobulins]. / Purpura rhumatoïde et immunoglobulines intraveineuses.

Henoch-Schönlein purpura is a vasculitis usually with a benign course. Abdominal symptoms occur in 70% of cases, with possible intussusception or intestinal perforation. There is no clear evidence of the efficacy of a treatment in complicated cases of Henoch-Schönlein purpura. Corticosteroids improve abdominal pain but they do not have any effect on renal involvement or prevention of relapses. Intravenous immunoglobulins have been efficient in some cases with recurrent abdominal symptoms or progressive renal lesions. We report the case of a 19-year-old patient with severe abdominal involvement and early renal manifestations of Henoch-Schönlein disease, rapid and sustained improvement was obtained by intravenous immunoglobulins given during 48 hours.

[MRI and CT of parotid diseases in HIV-positive patients]. / IRM-et TDM de la pathologie parotidienne chez le patient VIH positif.

We report five cases of HIV patients with parotid pathology during a period of one year. All patients (4 men and 1 woman between the ages of 32 and 47 years) had a MRI or a CT confirmation of the parotid gland lesion. Three patients exhibited parotid gland cysts, one a nonspecific chronic inflammation and one an enlargement of the parotid gland with some benign lymphoepithelial lesions. Two patients underwent ultrasound-guided fine-needle aspiration biopsy. The cytology examination confirmed the benignity of the lesions. Finally, we expose the clinical management of parotid enlargements in HIV-positive patients.

Safety and immunogenicity of Haemophilus influenzae type b conjugate vaccine (PedvaxHIB) in Papua New Guinean children.

BACKGROUND: In view of high mortality and morbidity from Haemophilus influenzae type b (Hib) in young Papua New Guinean children, the incorporation of a Hib conjugate vaccine into a nationwide immunization program would be of major public health benefit. The choice of the Hib conjugate vaccine will be based on the evaluation of several Hib conjugate vaccines, after consideration of such factors as the ease of incorporation into the current vaccination schedule, cost, kinetics of antibody responses and safety. METHODS: This study evaluated the safety and immunogenicity of Hib polysaccharide-Neisseria meningitidis outer membrane protein complex conjugate vaccine (PRP-OMPC) in Papua New Guinea. 95 children were recruited at Goroka Base Hospital, Eastern Highlands Province, and enrolled in the study. PRP-OMPC was administered at ages 2, 4 and 12 to 15 months. Blood was collected before each dose, one month after the second and booster doses, and at ages 18 and 24 months. Antibody to PRP (anti-PRP) was measured by radioimmunoassay. RESULTS: PRP-OMPC was generally well tolerated. At successive sampling times from the prevaccination bleed through the 1-month post-booster bleed, geometric mean titres were 0.18, 1.45, 2.54, 1.03 and 8.05 micrograms/ml, respectively (n = 60). The proportions of subjects with anti-PRP titres > or = 1.0 microgram/ml were 2%, 62%, 73%, 47% and 93%, respectively (n = 60). Persistence of anti-PRP was ascertained in 41 subjects. The GMTs at 18 and 24 months were 3.42 and 2.0 micrograms/ml, respectively. CONCLUSIONS: PRP-OMPC was found to be immunogenic after the first dose and to elicit a robust booster response. Antibody titres persisted until age 24 months, at which time 100% of subjects had anti-PRP > or = 0.15 microgram/ml. These results are consistent with previous studies in US Native American infants and in Gambian infants.

Regional differences in the risk of triple class failure in European patients starting combination antiretroviral therapy after 1 January 1999.

Regional differences across Europe in triple class failure (TCF; the failure of each of the three separate main classes of antiretrovirals (ARVs) with a viral load >1000 HIV-1 RNA copies/mL for >4 months) have not been described. A total of 1956 patients started combination ARV therapy after 1 January 1999, of whom 123 patients developed TCF [6.3%; incidence 16.7 per 1000 person-years of follow-up; 95% confidence interval (CI) 13.7-19.6]. After adjustment, patients from Eastern Europe had a significantly increased incidence of TCF compared with patients from Southern Europe/Argentina (3.05; 95% CI 1.36-6.82; P=0.0067) while patients taking either a boosted protease inhibitor regimen (0.33; 95% CI 0.15-0.74; P=0.0072) or a nonnucleotide reverse transcriptase inhibitor (NNRTI)-based regimen (0.59; 95% CI 0.37-0.94; P=0.026) had a reduced incidence of TCF.

[Febrile status and exanthema following a trip to the jungle]. / Status febrilis und Exanthem nach Dschungelaufenthalt.

A 30-year-old female entered the emergency room for medical advice because of progressive deterioration of general health with headache, arthralgias, myalgias and fever after a vacation of three weeks in Malaysia and Hong Kong. Because of persistent fever, lymphadenopathy, slight leuco- and thrombocytopenia and only insignificantly elevated humoral signs of an inflammatory process, the patient was treated symptomatically after exclusion of malaria. A viral disease was suspected. Two days later, an exanthema erupted suddenly on the trunk. Pinhead-sized livid, flat macules, increasing in size within hours and spreading to the extremities, were observed. Further investigations revealed a significantly elevated titer of IgG directed against rickettsia conorii, leading to the diagnosis of Mediterranean spotted fever. Under antibiotic treatment with tetracycline, the aforementioned findings regressed within few days, and the patient recovered completely.

Benzothiadiazole induces disease resistance in Arabidopsis by activation of the systemic acquired resistance signal transduction pathway.

Benzothiadiazole (BTH) is a novel chemical activator of disease resistance in tobacco, wheat and other important agricultural plants. In this report, it is shown that BTH works by activating SAR in Arabidopsis thaliana. BTH-treated plants were resistant to infection by turnip crinkle virus, Pseudomonas syringae pv 'tomato' DC3000 and Peronospora parasitica. Chemical treatment induced accumulation of mRNAs from the SAR-associated genes, PR-1, PR-2 and PR-5. BTH treatment induced both PR-1 mRNA accumulation and resistance against P. parasitica in the ethylene response mutants, etr1 and ein2, and in the methyl jasmonate-insensitive mutant, jar1, suggesting that BTH action is independent of these plant hormones. BTH treatment also induced both PR-1 mRNA accumulation and P. parasitica resistance in transgenic Arabidopsis plants expressing the nahG gene, suggesting that BTH action does not require salicylic acid accumulation. However, because BTH-treatment failed to induce either PR-1 mRNA accumulation or P. parasitica resistance in the non-inducible immunity mutant, nim1, it appears that BTH activates the SAR signal transduction pathway.

Down-regulation of hepatic and renal 11 beta-hydroxysteroid dehydrogenase in rats with liver cirrhosis.

BACKGROUND & AIMS: 11 beta-Hydroxysteroid dehydrogenase (11 beta-OHSD) enzymes are responsible for the interconversion of active 11 beta-hydroxycorticosteroids into inactive 11-ketoglucocorticosteroids and by that mechanism regulate the intracellular access of the steroids to the cognate receptor. A down-regulation of the shuttle of active to inactive glucocorticoids enhances access of glucocorticosteroids to both the glucocorticoid and the mineralocorticoid receptors. In liver cirrhosis, enhanced mineralocorticoid and glucocorticoid effects are observed. We therefore investigated the impact of liver cirrhosis after bile duct ligation on the transcription and activity of 11 beta-OHSD1 and 11 beta-OHSD2 in the corresponding tissues. METHODS: Messenger RNA from 11 beta-OHSD1 and 11 beta-OHSD2 was assessed by reverse-transcription polymerase chain reaction; activity was assessed by measuring the interconversion of corticosterone to dehydrocorticosterone. The effect of bile and bile salts was determined using COS-1 cells transfected with 11 beta-OHSD1 or 11 beta-OHSD2. RESULTS: In liver tissue, the messenger RNA ratios of 11 beta-OHSD1 to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels and, in kidney tissue, the ratios of 11 beta-OHSD2 to GAPDH levels decreased after induction of liver cirrhosis. The 11 beta-OHSD activities were correspondingly reduced. Bile and individual bile salts inhibited 11 beta-OHSD1 and 11 beta-OHSD2 oxidative activity in transfected COS-1 cells. CONCLUSIONS: These findings indicate that in liver cirrhosis the mineralocorticoid and glucocorticoid receptor-protecting effects by the 11 beta-OHSD isoenzymes are down-regulated and that by the same mechanism the glucocorticoid and mineralocorticoid effects are enhanced.

Benzothiadiazole, a novel class of inducers of systemic acquired resistance, activates gene expression and disease resistance in wheat.

Systemic acquired resistance is an important component of the disease resistance repertoire of plants. In this study, a novel synthetic chemical, benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH), was shown to induce acquired resistance in wheat. BTH protected wheat systemically against powdery mildew infection by affecting multiple steps in the life cycle of the pathogen. The onset of resistance was accompanied by the induction of a number of newly described wheat chemically induced (WCI) genes, including genes encoding a lipoxygenase and a sulfur-rich protein. With respect to both timing and effectiveness, a tight correlation existed between the onset of resistance and the induction of the WCI genes. Compared with other plant activators, such as 2,6-dichloroisonicotinic acid and salicylic acid, BTH was the most potent inducer of both resistance and gene induction. BTH is being developed commercially as a novel type of plant protection compound that works by inducing the plant's inherent disease resistance mechanisms.

Simple DNA extraction method for dried blood spots and comparison of two PCR assays for diagnosis of vertical human immunodeficiency virus type 1 transmission in Rwanda.

Dried blood spots (DBS) on filter paper facilitate the collection, transport, and storage of blood samples for laboratory use. A rapid and simple DNA extraction procedure from DBS was developed and evaluated for the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection in children by an in-house nested-PCR assay on three genome regions and by the Amplicor HIV-1 DNA prototype assay version 1.5 (Roche Molecular Systems). A total of 150 samples from children born to HIV-1-infected mothers were collected in Kigali, Rwanda, in parallel as DBS and as peripheral blood mononuclear cell (PBMC) pellets. The results obtained on DBS by the two PCR assays were compared to the results of nested PCR on PBMCs. Of 150 PBMC samples, 10 were positive, 117 were negative, and 23 were indeterminate for HIV-1 infection. In DNA extracted from filter papers and amplified by using the in-house nested PCR, 9 of these 10 positive samples (90%) were found to be positive, and 1 was found to be indeterminate (only the pol region could be amplified). All of the negative samples and all of the 23 indeterminate samples tested negative for HIV-1 infection. When we used the Amplicor DNA test on DBS, all of the 10 PBMC-positive samples were found to be positive and all of the 23 indeterminate samples were found to be negative. Of the PBMC-negative samples, 115 were found to be negative and 2 were found to be indeterminate. We conclude that this simple rapid DNA extraction method on DBS in combination with both detection methods gave a reliable molecular diagnosis of HIV-1 infection in children born to HIV-infected mothers.