Novel role of fluorescent in situ hybridization technique (FISH) in recommended surgical margins of dermatofibrosarcoma protuberans: A preliminary study.

BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is a rare skin tumor. There is no standard recommendation for its surgical management. The currently used histological analysis are HES (hematoxylin eosin saffron) staining and immunohistochemistry for CD34 expression in particular cases. Fluorescent in situ hybridization (FISH) technique is only used to qualify the DFSP as translocated or non-translocated and is not used as a diagnostic method. The aim of our study was to determine by FISH (as a diagnostic method) whether cancerous cells that could not be identified through HES staining±immunohistochemistry were present at the two-centimeter margins that were found to be tumor-free. METHODS: Samples from patients who underwent surgery between 2010 and 2018 were collected. Intralesional and peripheral (at 2cm margins) paraffin slides were included. An average of 7.4 slides per specimen was analyzed. Firstly, the preselected slides were reread by a senior pathologist to confirm the absence of microscopic findings of DFSP at 2cm margins. Secondly a FISH analysis was used as a quantitative diagnostic approach, in order to find the t(17;22) translocation. RESULTS: Among the seven specimens that included 2cm margins, two samples presented one or more translocations, which were not visible in standard morphology assessments at two centimeters tumor-free margins. CONCLUSIONS: FISH analysis can have a new role in defining tumor-free margins. This would reduce the incidence of disease recurrence after resection and improve the post-operative complementary care.

The infragluteal fold: An appraisal by MRI combined with an anatomic study.

PURPOSE: The gluteal region is a key element of beauty balance and sexual appearance. However, there is no clear anatomical description of the infragluteal fold, nor any classification exists allowing standardizing treatment of this area in case of jeopardisation. The purpose of this study was to perform an anatomical description of the infragluteal fold (IGF) matching radiological and anatomical findings in describing specifically raise of the fibrous component at the bone level. METHODS: Six volunteers (three males and three females) underwent an MRI scan (Siemens Aera® 1.5 T) of the pelvic region. T1 Vibe Morpho T2, Sag Space 3D, and Millimetric slices were performed in order to obtain a more detailed selection of the gluteal landmark. Trabecular connective tissue of the region was analyzed using Horos® ROI (region of interest) segmentation function. Four fresh cadavers (two males, two females, accounting for 8 hemipelvis) were dissected in order to compare the radiological findings. RESULTS: The infragluteal fold is a connectival fibrous band extending from the ramus of the ischium (but not involving the ischial tuberosity, for a length of 21 mm ± 2 and 21 mm ± 3), the apex of the sacrum (for a length of 13 ± 2 and 11 mm ± 2), and the coccyx (for a length of 19 mm ± 2 and 20 mm ± 2, all measures referring to volunteers and cadavers, respectively) reaching superficially the dermis of the medial one-third of the cutaneous fold. No significant difference was found between volunteer and cadaver group in MRI measurement of bony origins, or between MRI and cadaveric dissection measurements. CONCLUSION: Knowledge of this structure will define novel surgical techniques in infragluteal fold restoration.

Coverage of radial forearm flap donor site with full thickness skin graft and Matriderm®: An alternative reliable solution?

PURPOSE: After harvesting a radial forearm flap (RFF) an optimal aesthetic and functional restitution of the donor site is required. In order to cover the secondary defect of the donor site, several solutions are currently available, but there is still no real evidence of the most reliable option. A retrospective study was conducted in order to evaluate a new technique of forearm coverage with artificial dermis: the association of full thickness skin graft (FTSG) with Matriderm®. METHODS: Our study included all RFF performed during a 34-month period. Forty-three forearm secondary defects after harvesting a RFF (16 men, 27 female) were included. Forearm donor site was covered using three techniques: a simple FTSG, split thickness skin graft (STSG) with Matriderm® or FTSG with Matriderm®. Clinical evaluations based on residual functionality, skin quality and aesthetic result were assessed using respectively the Disabilities of the Arm, Shoulder and Hand (DASH) score, the Vancouver Scar Scale (VSS) score and a blind panel questionnaire. RESULTS: FTSG with Matriderm® showed an improved DASH (10.6/100) and VSS score (5.5/13) if compared to the other techniques, mean surgeon satisfaction score was 3/5, mean patient satisfaction score was 3/5 in the FTSG with Matriderm® group. CONCLUSION: The results of this study revealed that the new association of FTSG with Matriderm® improves the DASH score and the aesthetic outcomes resulting to be a reliable solution in treating full thickness forearm skin defects after RFF harvesting.

Fluorescent indocyanine green angiography: Preliminary results in microsurgery monitoring.

INTRODUCTION: Pedicled flaps and free-tissue transfer flaps are used routinely to reconstruct hard and soft tissue defects in head and neck, limb, hand, thoracic and abdominopelvic reconstructive surgery. But failure remains a constant concern, particularly in free-tissue transfers. Usually failure is due to blood supply compromise. Indocyanine green (ICG), a fluorescent dye is a suitable tracer for vessel perfusion. The objective of this study is to evaluate the fluorescent indocyanine green angiography (FA ICG) in free flaps procedures. MATERIEL AND METHODS: Patients who had microsurgical flap reconstruction were included during the study period in a single center. The FA ICG was used at specific times. Intra-veinous injections of 0.1mg/kg of INFRACYANINE® (concentration 2.5mg/mL) were done intraoperatively. The Fluobeam® device programmed on sensitivity and mapping to interpret the data, was used. These different injections allowed to checked skin paddle perforators vessels, osseous perforators vessels, arterial and venous patency after anastomosis and the cutaneous, muscular and osseous perfusion. RESULTS: A total of 12 patients enrolled were 10 males and 1 female. Their mean age was 54.5 years (range 25-75 years). Of the 12 flaps, 8 were free flaps with 4 fibular flaps (3 for mandibular reconstruction and one for femur reconstruction); 2 radial forearm flaps for maxillary reconstruction; one latissimus free flap for tibia skin coverage and one retroauricular fasciocutaneous free flap for thumb skin coverage. We got to modify specific steps during surgery with 8 patients by using the FA ICG to anticipate potential complications: modifying the draw of the skin paddle, recut of this paddle, modifying the osteotomies, re-doing the anastomosis or modifying the position of the pivot point. DISCUSSION: Evaluation of microvascular flap perfusion is still based on subjective clinical features. Clinical monitoring is observer-dependent and does not allow information sharing, test reproducibility, and consistent postoperative follow-up. The successful of salvage rate is linked to the delay between the onset of ischemia and its clinical assessment. FA ICG could be a reliable method for monitoring free-tissue transfers. This technique is objective, non invasive and facilitate a complex reconstructive procedure to augment is liability. This technique may be used such a pedagogical tool for young practitioners in their first microsurgery procedures.

Latissimus dorsi breast reconstruction with or without implants: A comparison between outcome and patient satisfaction.

BACKGROUND: The latissimus dorsi (LD) flap is largely used in breast reconstruction. The aim of this study is to evaluate and compare the advantage of the LD flap in association with an implant to that of LD flap in association with lipofilling to add additional volume following breast reconstruction by assessing the number of revision procedures, length of hospitalization, complication rate, and patient satisfaction. METHODS: A retrospective cohort study was performed to evaluate complication rate, number and type of revision procedures required, hospitalization time, and overall duration of the reconstructive process in patients undergoing LD breast reconstruction in our department between January 2012 and March 2015. Patient satisfaction was assessed using BREAST-Q®. RESULTS: Ninety-five breast reconstructions performed on 90 patients were included. In 38 patients, 42 breasts were reconstructed using LD flap in association with lipofilling (Group 1). In the remaining 52 patients, 53 breast reconstructions were performed using LD flap in association with an implant (Group 2). Findings concerning total hospitalization time, overall duration of the reconstructive process, and the distribution of supplementary surgical procedures demonstrated no statistically significant differences between the two groups. However, the surgical complication rate was higher in Group 2 than in Group 1 (14.2% vs. 18.8%, respectively). Regarding BREAST-Q, a detailed analysis of the "satisfaction with breasts" domain showed a higher score for Group 1 than for Group 2. CONCLUSIONS: The association of a breast implant with LD reconstruction does not decrease breast reconstruction time in terms of the number of revision procedures and hospitalization time, exposes patients to a higher complication rate, and does not improve patient satisfaction.

Postoperative complications of male to female sex reassignment surgery: A 10-year French retrospective study.

In primary male to female (MTF) sex reassignment surgery (SRS), the most frequent postoperative functional complications using the penoscrotal skin technique remain neovaginal stenosis, urinary meatal stenosis and secondary revision surgery. We aimed to retrospectively analyze postoperative functional and anatomical complications, as well as secondary procedures required after MTF SRS by penile skin inversion. All patients operated on for MTF SRS, using the inverted technique, from June 2006 to July 2016, were retrospectively reviewed. The minimum follow-up was one year (five-years maximum follow-up). Soft postoperative dilationprotocol was prescribed until complete healing of the vagina. We did not prescribe long-term hard dilation systematically. Possible short-depth neovaginas were primarily treated with further temporary dilation using a hard bougie. Among the 189 included patients, we reported a 2.6% of rectovaginal wall perforations. In 37% of patients we had repeated compressive dressings and 15% of them required blood transfusions. Eighteen percent of patients presented with hematoma and 27% with early infectious complications. Delayed short-depth neovagina occurred in 21% of patients, requiring additional hard dilatation, with a 95.5% success rate. Total secondary vaginoplasty rate was 6.3% (4.7% skin graft and 3.7% bowel plasty). Secondary functional meatoplasty occurred in 1% of cases. Other secondary cosmetic surgery rates ranged between 3 to 20%. A low rate of secondary functional meatoplasty was showed after MTF SRS by penile skin inversion. Hard dilation was prescribed in case of healed short-depth vagina, with good efficiency in most of cases. Secondary vaginoplasty was required in cases of neovagina stenosis or persisting short-depth neovagina after failure of hard dilation protocol.

Growth and adhesion to HT-29 cells inhibition of Gram-negatives by Bifidobacterium longum BB536 e Lactobacillus rhamnosus HN001 alone and in combination.

OBJECTIVE: The aim of this study was to test the inhibitory effect of supernatants of broth cultures of Bifidobacterium longum BB536 and Lactobacillus rhamnosus HN001, both individually and in combination, against Gram-negative strains (uropathogens, enteropathogens and a reference strain). Moreover, in vitro protection of B. longum BB536 and L. rhamnosus HN001, both individually and in combination, against pathogen adhesion to HT-29 cell line, was investigated. MATERIALS AND METHODS: The inhibitory activity was performed by the agar diffusion test and in vitro antagonistic activity against pathogen adhesion to human epithelial intestinal HT-29 cells was performed using standardized culture techniques. RESULTS: The study showed that B. longum BB536 and L. rhamnosus HN001, individually and in combination have inhibitory activity against the majority of the Gram negative strains tested. Furthermore, the results showed that both probiotic strains have a good capacity to inhibit pathogenic adhesion to HT-29 cells. Moreover, the ability of B. longum BB536 and L. rhamnosus HN001 to inhibit pathogenic adhesion increased when they were used in combination. DISCUSSION: The combination of B. longum BB536 and L. rhamnosus HN001 showed inhibitory activity against Gram-negatives and an improved ability to reduce their adhesion properties and to compete with them. CONCLUSIONS: The simultaneous presence of the two-probiotic strains could promote competitive mechanisms able to reduce the adhesion properties of pathogen strains and have an important ecological role within the highly competitive environment of the human gut.

A NOVEL MODEL TO EVALUATE THE LEARNING CURVE IN MICROSURGERY: SERIAL ANASTOMOSIS OF THE RAT FEMORAL ARTERY.

The rat's femoral artery is definitely the most frequently used model in microsurgical training for its easy dissection. Our model, consisting in the creation of several anastomoses in a row, helps the novice surgeon to assess his microsurgical level and to improve his capacity. Indeed, this leads to an amplification of the trainee surgeon's mistakes, which add up to each other as the anastomoses are performed. We propose a simple method to evaluate the surgeon microsurgical skills during the training.

Microbiological characteristics of the probiotic strains B. longum BB536 and L. rhamnosus HN001 used in combination.

AIM: The aim of this study was to investigate the antagonistic activity between the probiotic strains Bifidobacterium longum (B. longum) BB536 and Lactobacillus rhamnosus (L. rhamnosus) HN001 (ZirCombi, Alfa Wassermann S.p.A., Italy) and to evaluate for the strains tested alone and in combination the resistance in simulated gastrointestinal conditions, the ability to adhere to epithelial intestinal cells and their competition for adhesion. METHODS: The antagonism between B. longum BB536 and L. rhamnosus HN001 was tested modifying the agar diffusion method. The in vitro resistance to gastrointestinal condition of the two strains used alone or in combination was tested using low pH (3.0, 2.5 and 2.0) and different concentrations of bile salts (0.3%, 0.5% and 0.7%). The adhesion ability and the competition for adhesivity on human colon cancer HT-29 cells were tested modifying quantitative methods described in literature. RESULTS: The results demonstrated that B. longum BB536 and L. rhamnosus HN001 showed no in vitro inhibition effect each other and a good resistance to low pH and to different concentrations of bile salts, that was enhanced when they were tested in combination. Moreover, the two strains tested alone and in combination showed a good adhesion on HT-29 cells and no mechanism of competition. CONCLUSION: The study suggests that B. longum BB536 and L. rhamnosus HN001 used in combination show no antagonism and could have functional endosymbiotic effects on intestinal host-microbiota.

Virological response rates for telaprevir-based hepatitis C triple therapy in patients with and without HIV coinfection.

OBJECTIVES: Pegylated-interferon/ribavirin dual therapy for hepatitis C virus (HCV) infection has a lower sustained virological response (SVR) rate in HIV/HCV-coinfected patients than in HCV monoinfected patients, but little is known about the relative effectiveness of teleprevir-based triple therapy in the two groups. METHODS: Data on 33 coinfected and 116 monoinfected patients were analysed on an intention-to-treat basis. SVR12 was defined as undetectable HCV RNA at week 12 post-end-of-treatment, severe anaemia as haemoglobin ≤ 89 g/L or a drop of ≥ 45 g/L, and advanced fibrosis/cirrhosis as Fib-4 ≥ 3.25. All coinfected patients had well controlled HIV infection. RESULTS: The groups were similar in age, gender, percentage with Fib-4 ≥ 3.25 and HCV viral load, but differed in previous treatment response, with more coinfected patients being nonresponders or treatment-intolerant (75.8% vs. 50.0% for monoinfected patients; P < 0.01). During treatment, the percentages of patients with undetectable HCV RNA were similar, but, surprisingly, this percentage tended to be higher in coinfected patients. SVR12 rates were 60.6% in coinfected patients vs. 42.2% in monoinfected patients (P = 0.06). In multivariable analysis, SVR12 was associated with HIV infection [odds ratio (OR) 3.55; P < 0.01], African American race (OR 0.37; P = 0.03) and previous treatment response (OR 0.46; P = 0.03). Rates of severe anaemia (45.5 vs. 58.6% in coinfected and monoinfected patients, respectively; P = 0.18) were similar in the two groups, but rash (15.2 vs. 34.5%, respectively; P = 0.03) and rectal symptoms (12.1 vs. 43.1%, respectively; P < 0.01) were less common in coinfected patients. CONCLUSIONS: Virological responses of coinfected and monoinfected patients did not differ significantly, but tended to be higher in coinfected patients, who had a 60.6% SVR12 rate. Telaprevir-based triple therapy is a promising option for coinfected patients with well-controlled HIV infection.

Activity of Melaleuca alternifolia (tea tree) oil on Influenza virus A/PR/8: study on the mechanism of action.

Our previous study demonstrated that Melaleuca alternifolia (tea tree) oil (TTO) had an interesting antiviral activity against Influenza A in MDCK cells. In fact, when we tested TTO and some of its components, we found that TTO had an inhibitory effect on influenza virus replication at doses below the cytotoxic dose; terpinen-4-ol, terpinolene, and alfa-terpineol were the main active components. The aim of this study was to investigate the mechanism of action of TTO and its active components against Influenza A/PR/8 virus subtype H1N1 in MDCK cells. None of the test compounds showed virucidal activity nor any protective action for the MDCK cells. Thus, the effect of TTO and its active components on different steps of the replicative cycle of influenza virus was studied by adding the test compounds at various times after infection. These experiments revealed that viral replication was significantly inhibited if TTO was added within 2h of infection, indicating an interference with an early step of the viral replicative cycle of influenza virus. The influence of the compound on the virus adsorption step, studied by the infective center assay, indicated that TTO did not interfere with cellular attachment of the virus. TTO did not inhibit influenza virus neuraminidase activity, as shown by the experiment measuring the amount of 4-methylumbelliferone, cleaved by the influenza virus neuraminidase from the fluorogenic substrate 2'-O-(4-methylumbelliferyl)-N-acetylneuraminic acid. The effect of TTO on acidification of cellular lysosomes was studied by vital staining with acridine orange using bafilomycin A1 as positive control. The treatment of cells with 0.01% (v/v) of TTO at 37°C for 4h before staining inhibited the acridine orange accumulation in acid cytoplasmic vesicles, indicating that TTO could inhibit viral uncoating by an interference with acidification of intralysosomal compartment.

Antipoliovirus activity and mechanism of action of 3-methylthio-5-phenyl-4-isothiazolecarbonitrile.

Our previous studies described the synthesis and the antiviral activity of 3,4,5-trisubstituted isothiazole derivatives that were found to be particularly effective against enteroviruses. Compound 3-methylthio-5-phenyl-4-isothiazolecarbonitrile (IS-2) exhibited an interesting anti-poliovirus activity with a high selectivity index. In the present study we investigated the mechanism of action of this compound. Studies on the time of IS-2 addition to poliovirus type 1 infected cells suggested that the compound may inhibit some early process of viral replication. In order to determine its mechanism of action, we evaluated the rate of attachment and internalization of purified [³H]uridine-labeled poliovirus to HEp-2 cells in the presence or absence of IS-2. No effect on poliovirus adsorption and internalization to host cells was detected. We also investigated the influence of the compound on virus uncoating using labeled poliovirus and measuring the radioactivity of oligoribonucleotides formed from viral RNA susceptible to ribonuclease. These experiments demonstrated that poliovirus uncoating is influenced by IS-2 action.

High frequency of Chlamydophila pneumoniae infections: patients with peripheral arterial disease and those with risk factors for cardiovascular diseases compared to normal subjects.

The role of bacterial infections, mainly Chlamydophila pneumoniae, on atherosclerotic processes as well as the therapeutic utility of additional antibiotic treatment is still an open question. In this study we compared the serological profiles of 160 patients (80 with peripheral arterial disease (PAD), diagnosed with an ankle/brachial index (ABI) ≤ 0.9 and 80 with risk factors for cardiovascular disease - CVD) with those of 80 healthy subjects, serum levels of specific C. pneumoniae antibodies using the microimmunofluorescence test. Our results show that PAD patients had a higher frequency of C. pneumoniae infection than those with risk factors for cardiovascular disease. This frequency was lower if compared to the previous two groups in controls. 44 out of the 80 (55%) patients with PAD and 34 out of the 80 (42.58%) subjects with risk factors for cardiovascular disease were seropositive while only 24 of the 80 (30%) healthy subjects showed seropositivity to C. pneumoniae. Furthermore, higher anticorpal titers were also found in patients with peripheral arterial disease and in patients with cardiovascular risk factors if compared to healthy subjects. On the basis of these results, we confirm that C. pneumoniae infection is frequent in peripheral arterial disease patients and we believe that it could be considered as an additional risk factor involved in the pathogenesis of this disease.

Review article: adherence to medication for chronic hepatitis C - building on the model of human immunodeficiency virus antiretroviral adherence research.

BACKGROUND: Treatment of hepatitis C virus (HCV) infection with pegylated interferon/ribavirin achieves sustained virological response in up to 56% of HCV mono-infected patients and 40% of HCV/human immunodeficiency virus (HIV)-co-infected patients. The relationship of patient adherence to outcome warrants study. AIM: To review comprehensively research on patient-missed doses to HCV treatment and discuss applicable research from adherence to HIV antiretroviral therapy. METHODS: Publications were identified by PubMed searches using the keywords: adherence, compliance, hepatitis C virus, interferon and ribavirin. RESULTS: The term 'non-adherence' differs in how it is used in the HCV from the HIV literature. In HCV, 'non-adherence' refers primarily to dose reductions by the clinician and early treatment discontinuation. In contrast, in HIV, 'non-adherence' refers primarily to patient-missed doses. Few data have been published on the rates of missed dose adherence to pegylated interferon/ribavirin and its relationship to virological response. CONCLUSIONS: As HCV treatment becomes more complex with new classes of agents, adherence will be increasingly important to treatment success as resistance mutations may develop with suboptimal dosing of HCV enzyme inhibitors. HIV adherence research can be applied to that on HCV to establish accurate methods to assess adherence, investigate determinants of non-adherence and develop strategies to optimize adherence.

Chronic peripheral arteriopathy is associated with seropositivity to Chlamydia pneumoniae.

The study was carried out to clarify the correlation between Chlamydia pneumoniae infection and peripheral arterial disease (PAD). The level of specific antibodies of the 133 consecutive patients suffering from PAD at 2nd stage of Leriche's classification were compared with 60 healthy controls by using a commercial Micro-IF Test. A higher incidence of serological evidence of C. pneumoniae infection was found in the patients (106/133) than in controls (6/60). These results are in agreement with other findings that measured the infection in atheromasic plaques. A strong cause-effect relationship between bacterial infection and peripheral arterial disease was not found, but the raised seropositivity level could be considered as a target for medical therapy of PAD.

Use of a microquantity enzyme immunoassay in a large-scale study of measles, mumps and rubella immunity in Italy.

The seroprevalence of antibodies to measles, mumps, and rubella viruses (MMR) was determined in 1498 subjects in Catania, Italy, ranging in age from 1 month to 25 years. The study population was divided into seven age groups and screened by enzyme immunoassay using microquantities (10 microl) of whole blood collected by fingerprick on filter paper discs. The results showed that seroconversion for measles (87.6%) and mumps (73.2%) occurred between 6 and 10 years of age. The seroprevalence of antibodies to rubella virus increased slowly through the age groups, reaching the highest rate (93.3%) between 16 and 20 years of age. Passively transmitted maternal antibodies to mumps and rubella were absent in babies between 5 and 8 months of age, and a few cases positive for measles antibodies were found among babies 6 and 7 months of age. The enzyme immunoassay was demonstrated to be suitable for low-cost large-scale screening for MMR immunity. The rate of vaccine failure was also evaluated and found to be 9.5% for the measles virus, 12.9% for the mumps virus and 0.0% for the rubella virus.

Improvement in establishing the period of rubella virus primary infection using a mild protein denaturant.

A more precise determination of the date of primary rubella infection in pregnancy is obtained by application of the urea denaturation method. Two denaturation buffers containing 6 and 8 M urea were compared for the detection of rubella IgG avidity by enzyme immunoassay. The results demonstrate that IgG avidity detected by 6M urea increases more rapidly, from 16% on the third day to 51% on the 46th day after the rash, than that obtained by 8 M urea, from 13 to 36%. The specific rubella IgG avidity detected by 6 M urea denaturation buffer increased significantly in paired sera obtained at an interval of 15 days. The same samples did not show any significant increase of IgG avidity when an 8 M urea denaturation buffer was used. The use of a mild denaturant such as 6 M urea indicates the presumptive date of primary rubella infection in pregnancy within 2 months of sampling.

Detection of immunoglobulin G to a Sindbis-related virus by a membrane antigen enzyme immunoassay.

We determined the seroprevalence of a Sindbis-related virus isolated for the first time in 1975 from ticks in south-east Sicily and typed by Gresikova et al. in 1978. An indirect enzyme immunoassay based on viral membrane antigen for coating microtiter strips was used for the detection of immunoglobulin G to the Sindbis-related virus. The method appeared more sensitive than a similar enzyme immunoassay based on crude lysate antigen. Comparison of the results obtained from sera tested both by membrane antigen enzyme immunoassay and microneutralization test showed 92% agreement, while the agreement between microneutralization test and crude antigen enzyme immunoassay was 76%. An overall elevated seroprevalence (63.66%) was found in a population group living in and around the area of first isolation and seroprevalence in different age groups was also studied.

Anti-HCMV serologic response in patients with acute viral hepatitis.

The authors point out a high serological incidence of HCMV infection in the course of HCV hepatitis compared to a low incidence detected in HBV and HAV hepatitis. Three immunoglobulin classes (IgG, IgM, and IgA) specific to HCMV were investigated. Antibody titers of each immunoglobulin class are reported for all the cases of HCMV infection.

Detection of immunoglobulin G to measles virus, rubella virus, and mumps virus in serum samples and in microquantities of whole blood dried on filter paper.

Immunity to measles virus, rubella virus, and mumps virus was determined by EIA in serum samples and in dried whole blood specimens spotted on Whatman filter paper (5 mm in diameter). Both specimens were obtained from each patient by venepuncture and finger prick. Ten microliters of whole blood is enough to detect antibodies to these three different viruses. The comparison of the results obtained by EIA from 227 serum and whole blood samples have demonstrated close agreement: 98.6% for measles virus, 99.1% for rubella virus, and 96.0% for mumps virus. Moreover, 96 whole blood samples can be tested in a microtiter plate and can be stored at room temperature for 15 days or at +4 degrees C for several months. Therefore, whole blood dried on filter paper is a convenient alternative method for collecting and transporting specimens, it is easier and safer than venepuncture, and could be used for large-scale epidemiological studies, especially in newborns. This method could solve the problem of sampling, especially in young children, and could simplify studies of vaccine efficacy.