María de Oña

Etiology of Sporadic Cases of Pediatric Acute Gastroenteritis in Asturias, Spain, and Genotyping and Characterization of Norovirus Strains Involved

ABSTRACT From November 2000 to October 2001, a reverse transcription-PCR using primers directed to the norovirus RNA polymerase coding region was included in a viral and bacterial routine screening to diagnose sporadic cases of acute gastroenteritis among children in Asturias, Spain. The role of noroviruses (8.6% of the positively diagnosed cases) as the cause of sporadic pediatric gastroenteritis was evaluated with respect to the detection rates of other gastroenteritis-associated viruses and bacteria. The results indicated that noroviruses were less common than rotaviruses (36.9%), Campylobacter spp. (28.8%), and Salmonella spp. (18.4%) but more frequent than astroviruses (4.3%), adenoviruses (3.8%), and Yersinia spp. (2.2%). Mixed infections involving noroviruses were rarely observed (0.5%). The presence of a norovirus-associated pediatric gastroenteritis peak in summer, as well as the complete absence of norovirus-associated cases in colder months, challenges the view that norovirus infections exclusively have wintertime seasonality. On the other hand, phylogenetic analysis of the amplified fragments showed that the norovirus strains responsible were closely related. A further study using the full-length capsid region showed that these strains could be included into genogroup II, Bristol/Lorsdale cluster, and were closely related to the 1995 and 1996 U.S. subset of strains associated with outbreaks recorded worldwide between 1995 and 1996.

Role of Metapneumovirus in Viral Respiratory Infections in Young Children

ABSTRACT The contribution of human metapneumovirus (hMPV) relative to that of other respiratory viruses as a cause of respiratory infections in children less than 1 year old has been evaluated. From October 2003 to April 2004, nasopharyngeal samples from 211 children less than 1 year old were analyzed to detect respiratory viruses. Respiratory syncytial virus (RSV) was the predominant virus isolated (96 children [45.5%]), followed by influenza A virus, parainfluenza virus, adenovirus, cytomegalovirus, and herpes simplex virus type 1, which were only occasionally detected. From January 2004 to April 2004, a nested retrotranscription-PCR, using in-house primers directed to the matrix protein gene of hMPV, was carried out on samples in which no other viruses were detected. hMPV was detected in 18 (16.2%) children, indicating that this virus was the second-most-frequent cause of viral respiratory infections in children less than 1 year old. The rate of hospitalization for RSV- and hMPV-infected children was higher than 75%. While RSV had a peak from December to February, hMPV was increasingly detected from January to April. The mean age of hMPV-infected children (6.44 ± 3.64 [mean ± standard deviation] months) was significantly higher than that of RSV-infected children (3.99 ± 2.96 [mean ± standard deviation] months). On the other hand, 64.3% of the RSV-infected children and 12.5% of the hMPV-infected children showed high levels of C-reactive protein. Although several authors have reported that clinical symptoms of hMPV-positive patients mirrored those of RSV-positive patients, differences between the two viruses can be found.

Human Papillomavirus Infection in a Male Population Attending a Sexually Transmitted Infection Service

Objective Human Papillomavirus (HPV) infection in men may produce cancer and other major disorders. Men play an important role in the transmission of the virus and act as a reservoir. The aim of this study was to determine the HPV-genotypes and their prevalence in a group of men attending a Sexually Transmitted Infection service. Patients and Samples Between July 2002 and June 2011, 1392 balanopreputial, 435 urethral, 123 anal, and 67 condyloma lesions from 1551 men with a mean age of 35.8±11.3 years old (range: 17–87) were collected for HPV-DNA testing. Methods A fragment of the L1-gene and a fragment of the E6/E7-genes were amplified by PCR. Positive samples were typed by hybridization. Results The HPV genome was detected in 36.9% (486/1318) balanopreputial and in 24.9% (101/405) urethral (p<0.0001) swabs from 38.1% (538) of 1469 men. Co-infections were present in 5.4% (80/1469) of cases. HPV was found in 43.9% (373/850) of men younger than 35 vs. 31.7% (187/589) of men aged >35. HPV was found in 59.4% (104) of 165 men with lesions (macroscopic or positive peniscopy), and in 22.8% (61/267) without clinical alterations. HPV was also detected in 71.4% (40/56) men with condylomata and in 58.7% (64/109) of men with positive peniscopy. Conclusions HPV prevalence in men was high and decreased with age. HPV was found more frequently in balanopreputial than in urethral swabs. There was a low rate of co-infections. Low-risk HPV vaccine genotypes were the most recurrent especially in younger. Although HPV has been associated with clinical alterations, it was also found in men without any clinical presentation. Inclusion of men in the national HPV vaccination program may reduce their burden of HPV-related disease and reduce transmission of the virus to non-vaccinated women.

Prevalence, evolution, and features of infection with human papillomavirus: A 15-year longitudinal study of routine screening of a women population in the north of Spain

Determination of the prevalence of type‐specific human papillomavirus (HPV) is important for the development of new vaccines and to prevent malignancy. The objective of this study was to determine HPV infection in two areas in the north of Spain, and their evolution in the last 15 years. Between 1991 and 2007, 7,930 fresh cervical swabs were obtained from 5,554 women (37.8 ± 11.8 years old). From them, 425 have been followed‐up for an average of 3.7 ± 2.08 years after sampling (range 2–14.6), and 71 for 7.7 ± 2.2 years (range 5–14). Methods based on polymerase chain reaction (PCR) were carried out. Samples from 1,598 (28.8%) women were positive for HPV: 40.9% were under 25 years of age, 34.2% in the 25–35 year age group, 27.2% in the 36–45 year age group, and 19.6% older than 45 years (P < 0.001). HPV was found in 34.4% of the women with cytological alterations versus 23% of women without cervical changes (P < 0.0001). HPV‐16 was present in 25.8% of the women, although the study identified 26 different HPV genotypes. After 3 years of follow‐up, HPV remained or became undetectable in 87% of the cases, and in 5 years 70.3%. The prevalence of HPV is associated with younger women and women with cytological changes in the cervix. Although HPV‐16 is more prevalent, HPV types not included in available vaccines were found the most commonly. The low 3‐year (even 5‐year) cumulative incidence rate of HPV infection suggests that cervical screening every 3 (or even 5) years is safe and effective. J. Med. Virol. 82:597–604, 2010.

Prevalence of HPV 16 and HPV 18 lineages in Galicia, Spain.

Genetic variants of human papillomavirus types 16 and 18 (HPV16/18) could differ in their cancer risk. We studied the prevalence and association with high-grade cervical lesions of different HPV16/18 variant lineages in a case-control study including 217 cases (cervical intraepithelial neoplasia grade 2 or grade 3 or worse: CIN2 or CIN3+) and 116 controls (no CIN2 or CIN3+ in two-year follow-up). HPV lineages were determined by sequencing the long control region (LCR) and the E6 gene. Phylogenetic analysis of HPV16 confirmed that isolates clustered into previously described lineages: A (260, 87.5%), B (4, 1.3%), C (8, 2.7%), and D (25, 8.4%). Lineage D/lineage A strains were, respectively, detected in 4/82 control patients, 19/126 CIN3+ cases (OR = 3.1, 95%CI: 1.0–12.9, p = 0.04), 6/1 glandular high-grade lesions (OR = 123, 95%CI: 9.7–5713.6, p<0.0001), and 4/5 invasive lesions (OR = 16.4, 95%CI: 2.2–113.7, p = 0.002). HPV18 clustered in lineages A (32, 88.9%) and B (4, 11.1%). Lineage B/lineage A strains were respectively detected in 1/23 control patients and 2/5 CIN3+ cases (OR = 9.2, 95%CI: 0.4–565.4, p = 0.12). In conclusion, lineages A of HPV16/18 were predominant in Spain. Lineage D of HPV16 was associated with increased risk for CIN3+, glandular high-grade lesions, and invasive lesions compared with lineage A. Lineage B of HPV18 may be associated with increased risk for CIN3+ compared with lineage A, but the association was not significant. Large well-designed studies are needed before the application of HPV lineage detection in clinical settings.

Noroviruses as cause of gastroenteritis in elderly patients

Background and aims: Gastroenteritis is one of the infections which are particularly important in elderly people. Knowledge of the main pathogens causing gastroenteritis in this group of patients, whose number will dramatically increase in coming decades, is essential. The contribution of group A rotavirus, adenovirus types 41 and 42, norovirus and astrovirus as causes of gastroenteritis among patients of all ages, especially those with over 65 was evaluated over an extended time period. Methods: A total of 4024 fecal samples, collected during seven seasons (October 2000 to September 2007), were tested with a commercial immunoassay (rotavirus) and an “in house” nested RT-PCR (adenovirus, norovirus and astrovirus). Results: Although norovirus was the second most common cause of gastroenteritis (7.9%) in the total population, it was predominant in the age group over the age of 6, causing 7.2% of gastroenteritis in the 6–16-year-old group, 8.6% in the 16–64-year-old group, and 11.1% in the >65-year group (p=0.001). In the last age group, norovirus was the most frequently detected (11.1%), followed by adenovirus (7.4%), astrovirus (3.6%) and rotavirus (3.3%) (p<0.0001). In addition, norovirus was rarely found in association with other viruses. Conclusions: Our data suggest that the elderly are highly vulnerable to certain infections, and indicate the need to introduce simple tests for an early identification of norovirus in cases of gastroenteritis affecting elderly patients, improving patient care by reducing unnecessary treatments and hospital stays.

Comparison of cytomegalovirus pp-65 antigenemia assay and plasma DNA correlation with the clinical outcome in transplant recipients

The relationship between quantitative antigenemia and plasma DNAemia was studied for monitoring cytomegalovirus (CMV) viremia in CMV infection (CMVI) or disease (CMVD), in 20 transplant recipients (13 CMD, seven CMVI). A total of 142 samples of blood were assayed for CMV‐DNA and pp‐65 antigenemia (CMV‐Ag). A quantitative correlation between both markers was found (P < 0.0001). First CMV antigenemia as well as first plasma DNA viral load was similar in CMVI and CMVD (29 vs. 24 CMV‐Ag+ cells/105 PMN; and 7445 vs. 12407 CMV‐DNA copies/ml). The maximum antigenemia was higher in CMVD than in CMVI (146 ± 87 vs. 61 ± 54 +cells/105 PMN, P < 0.05), but the highest CMV plasma viral load was similar in both groups (62592 ± 33000 vs. 42055 ± 38600 copies/ml). In nine patients, maximum antigenemia coincided with highest plasma DNA viral load, but in 10 highest DNAemia occurred 6 days later. On the contrary, antigenemia declined faster than CMV‐DNAnemia, after treatment.

Las infecciones por virus herpes simplex, Epstein-Barr, varicela zoster, papiloma humano, citomegalovirus o adenovirus no tienen relación con los adenocarcinomas nasosinusales☆

Objetivo Se ha relacionado a diversos virus con el desarrollo de tumores epidermoides de cabeza y cuello. No obstante, no existen estudios previos que relacionen a los adenocarcinomas nasosinusales (ACN) con la presencia de virus. El objetivo de este estudio es determinar, en una serie de ACN, la presencia de virus que se sabe desempenan un papel en el cancer. Material y metodo Se estudio mediante PCR 37 ACN, para determinar la presencia de ADN de virus de papiloma humano, virus de Epstein-Barr (VEB), virus herpes simplex, virus de la varicela zoster, adenovirus y citomegalovirus. Resultados Se detecto ADN de VEB en 3 (8,1 %) de las 37 muestras tumorales y ADN de citomegalovirus en 1 (2,7 %) de los 37 casos analizados. Conclusiones Nuestros resultados indican que los virus estudiados no desempenan papel alguno en la etiologia de los ACN.

Detection and quantification of Merkel cell polyomavirus. Analysis of Merkel cell carcinoma cases from 1977 to 2015

This study investigates the presence of Merkel cell polyomavirus (MCPyV) in skin lesions of patients with Merkel cell carcinoma (MCC). MCPyV was quantified using quantitative Real-Time-PCR (qRT-PCR) in 34 paraffinized MCC samples (resected/biopsied) originally taken between 1977 and 2015, and six non-MCC samples. In 31 (91.2%) MCC-individuals, MCPyV was detected. No virus was observed in any non-MCC tumor. Average age at diagnosis was 78.2 ± 9.35 (55-97) years for women (n = 19) and 69.5 ± 14.7 (45-91) for men (n = 15) (P = 0.04). MCC tumor location, known in 25 cases, was: 11 (44%) in the head region, 6 (24%) in upper limbs, 4 (16%) in lower limbs, and 4 (16%) in the trunk. All but one patient had received some sort of treatment: 15 (45.45%) underwent both radio and chemotherapy, 13 (39.39%) only surgery, 2 (6.06%) surgery, plus radio and chemotherapy, 2 (6.06%) surgery and chemotherapy, and 1 (3.03%) only radiotherapy. Follow up data were available for 21/34 patients: recurrence was recorded for 4 (19.04%), and metastasis for 13 (61.9%). Recorded data showed that 10 men and 5 women (total 44.1%) died during follow up, 7 (46.7%) of them within 2 years of diagnosis. Viral load was 5.8 ± 1.4 log copies/105 cells (3.1-8.6), independent of any variable. MCPyV was very frequent in MCC. It was principally associated with head and limb tumors, it more commonly affected men, who in this study were, on average, younger than women, and had high rates of recurrence and mortality. The amplification techniques described here are easily applied and suitable for detecting the presence of MCPyV virus in MCC.

Molecular Diagnosis of Human Papillomavirus Infections

Human Papillomavirus (HPV) is arguably the most common sexually transmitted agent worldwide, either in its clinical (genital warts) or subclinical presentation in men and women. The main interest in HPV relates to its recognized as a causal and necessary factor for cervical cancer one of the most common cancers in women (80% of cases in most developing countries, with an annual incidence of almost half a millon and a mortality rate of approximately 50%) [1-5], and other types of cancer, such as penis, anal or oral cancer [6].