Design of a data set of qPCR primers for the early region of Human Papillomavirus oncogenic types 16 and 18.

High-Risk Human Papillomavirus (HR-HPV) types 16 and 18 are estimated to be responsible for 72.4% of all HPV-related cancers worldwide in both men and women, including cervical, anal, penile, vulval, vaginal and head and neck cancers [1]. Important efforts worldwide have devoted to the study of these genotypes, throughout epidemiology and basic science approaches. Of particular interest are the genes from the early region (E), coding non-structural proteins. Early genes E1 and E2 products are involved in replication and transcription regulation, while E6 and E7 proteins are recognised for their oncogenic potential. In this data report, we described a set of primers based on reference sequences from HPV16 and HPV18 designed to cover the early region of these oncogenic genotypes. The design was based on multiple sequences alignment to identify the less conserved regions along the open reading frames (ORFs) E6, E7, E1 and E2. The design allows a highly stringent real time PCR essay ranged from 123 to 598 bp overlapping products for HPV16 (12 products in total) and from 183 to 526 bp for HPV18 (11 products in total), both spanning the early genomic region. The high annealing temperatures (Ta) and regions selected for primer bind were intended for genotypic specificity, without compromising the qPCR amplification efficiency (≥ 90%). Evaluation of qPCR conditions for primer set was performed using DNA standards as controls, generated from the HPV16 and 18 genomes clones. This provides relevant information for further multiple quantitative real-time PCR analysis (qPCR), using the SYBR green chemistry, which is is more affordable than generating multiple fluorescently labeled probes.

Human Papillomavirus Genotypes Infecting the Anal Canal and Cervix in HIV+ Men and Women, Anal Cytology, and Risk Factors for Anal Infection.

The incidence of anal intraepithelial neoplasias associated with HPV is rising worldwide. In the general population, this pathology is rare, but individuals living with HIV/AIDS are at a significantly higher risk. We aimed to study HPV infection and performed cytological screening to study the epidemiological and behavioral determinants in a group of men and women living with HIV from a region in Mexico with high HIV incidence. This was a cross-sectional study including adults living with HIV/AIDS performed in Merida (Mexico). We invited patients of public HIV/STD clinics and those affiliated with social organizations of people living with HIV to participate in the study. Participants responded to an instrument to assess their risky behaviors and clinical history. Swabs from the anal canal and cervix and anal cytology specimens were obtained by medical staff from women and by self-sampling from men. For the 200 participants, 169 men and 31 women, anal HPV PCR tests resulted in 59.8% positivity (62.6% of men and 45.2% of women), and 17 genotypes were identified. The most frequent high-risk (HR) types for the anal canal were: HPV33 (35.3%), HPV58 (20.6%), HPV66 (18.6%), HPV45 (17.6%), and HPV16 (14.7%). Multiple genotypes were found in over 80% of the participants. Receptive anal intercourse in the previous 12 months, inconsistent condom use, and detectable HIV titers (≥50 cc/mL) were associated with HPV infection (p < 0.05). Cytology (smears and liquid-based) identified that 34.6% of the participants had low-grade squamous intraepithelial lesions (LSILs), and 3.5% had high-grade squamous intraepithelial lesions (HSILs). Neither HPV nor lesions were associated with low CD4+ counts (<200 cells/mm3, p > 0.05). Of the women, 60% were infected in the cervix and 45% in the anal canal, with an agreement of at least one genotype in 90%. The HR-HPV types associated with HSILs were HPV66, 33, 52, 51, 45, 18, and 68.

Human IgG antibody responses to severe acute respiratory syndrome coronavirus 2 viral antigens receptor-binding domain, spike, and nucleocapsid, in vaccinated adults from Merida, Mexico.

Several vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been approved for controlling the coronavirus disease 2019 (COVID-19) pandemic worldwide. Antibody response is essential to understand the immune response to different viral targets after vaccination with different vaccine platforms. Thus, the main aim of this study was to describe how vaccination with two distinct SARS-CoV-2 vaccine preparations elicit IgG antibody specific responses against two antigenically relevant SARS-CoV-2 viral proteins: the receptor-binding domain (RBD) and the full-length spike (S). To do so, SARS-CoV-2 protein specific in-house enzyme-linked immunosorbent assays (ELISAs) were standardized and tested against serum samples collected from 89 adults, recipients of either a single-dose of the Spike-encoding mRNA-based Pfizer/BioNTech (Pf-BNT) (70%, 62/89) or the Spike-encoding-Adenovirus-5-based CanSino Biologics Inc. (CSBIO) (30%, 27/89) in Merida, Mexico. Overall, we identified an IgG seroconversion rate of 88% (68/78) in all vaccinees after more than 25 days post-vaccination (dpv). Anti-RBD IgG-specific responses ranged from 90% (46/51) in the Pf-BNT vaccine at 25 dpv to 74% (20/27) in the CSBIO vaccine at 42 dpv. Compared to the S, the RBD IgG reactivity was significantly higher in both Pf-BNT (p < 0.004) and CSBIO (p < 0.003) vaccinees. Interestingly, in more than 50% of vaccine recipients, with no history of COVID-19 infection, antibodies against the nucleocapsid (N) protein were detected. Thus, participants were grouped either as naïve or pre-exposed vaccinees. Seroconversion rates after 25 and more dpv varies between 100% in Pf-BNT (22/22) and 75% (9/12) in CSBIO pre-exposed vaccinees, and 89% (26/29) and 73% (11/15) in Pf-BNT and CSBIO naïve vaccine recipients, respectively. In summary, observed seroconversion rates varied depending on the type of vaccine, previous infection with SARS-CoV-2, and the target viral antigen. Our results indicate that both vaccine preparations can induce detectable levels of IgG against the RBD or Spike in both naïve and SARS-CoV-2 pre-exposed vaccinees. Our study provides valuable and novel information about the serodiagnosis and the antibody response to vaccines in Mexico.

[Molecular detection of human papillomavirus among patients with benign prostatic hyperplasia and prostate cáncer.] / Detección molecular del virus de papiloma humano en pacientes con hiperplasia prostática benigna y cáncer de próstata.

OBJECTIVE: To analyze the presenceof human papillomavirus in prostate and its associationwith prostate cancer. METHODS: A case-control study was conducted.Tissue samples with benign hyperplasia and prostatecancer were collected. Risk factors related to prostatecancer and human papillomavirus were assessedby a medical interview. Prostate tissue was obtainedby transrectal biopsy or transurethral resection. Theidentification of viral genome was assessed by the amplificationof 450 pb., from L1 gene. Real time PCR wasused to identified HPV genotypes 16 and 18. For dataanalysis, the χ2 test, Student's T test or Mann-WhitneyU test and OR were computed. RESULTS: Thirty and 99 with benign prostatehyperplasia were included in a 1:3 ratio, with a meanage of 69.44±9.22 years. The global prevalence of humanpapillomavirus was 15.2% being similar in bothcases (15.6%) and controls (15.1%) with no significantdifference (p = 0.572). Forty percent of the infectionswere persistent. From all positive samples, only in the40% were identified some of the genotypes analyzed(16 and 18). The group of patients with Gleason scorede > 7 had a virus prevalence of 16%. CONCLUSIONS: The results show the presence ofthe human papillomavirus genome in prostate tissuewith and without neoplasia; no association was foundbetween infection and prostate cancer.

OBJETIVOS: Analizar la presencia delvirus de papiloma humano en próstata y su asociacióncon cáncer.MATERIAL Y MÉTODOS: Se realizó un estudiode casos y controles, para lo cual se colectaronmuestras de tejido con hiperplasia benigna y concáncer de próstata. Se realizó una historia clínicapara conocer la presencia de los factores de riesgoasociados al cáncer de próstata, así como los relacionadoscon el virus. El tejido prostático fue obtenidopor biopsia transrectal o resección transuretral.La identificación del genoma viral se realizó amplificandoun fragmento de 450 del gen L1 por mediode una PCR clásica. Para la identificación de los genotipos16 y 18, se utilizó PCR tiempo real. Para elanálisis de los datos se utilizó la prueba de χ2, pruebade T de Student o U de Mann-Whitney y cálculode OR. RESULTADOS: Se incluyeron 32 pacientes concáncer de próstata y 99 con hiperplasia benigna depróstata en una relación 1:3. La media de edad fuede 69.44±9.22 años. La prevalencia global del virusde papiloma humano fue de 15.2% siendo similar en los casos y entre casos (15.6%) y controles (15.1%),no existiendo diferencia significativa (p=0.572). El40% de las infecciones eran persistentes. Del totalde las muestras positivas, solamente en el 40% seencontró alguno de los dos genotipos analizados (16y 18). Los pacientes con un puntaje de Gleason > 7tuvieron una prevalencia del virus de 16%. CONCLUSIONES: Los resultados ponen de manifiestola presencia del genoma del virus del papilomahumano en próstata con y sin neoplasia, no se encontróasociación entre la infección y el cáncer de próstata.

Detección molecular del Virus de Papiloma Humano en pacientes con hiperplasia prostática benigna y cáncer de próstata / Molecular detection of human papillomavirus among patients with benign prostatic hyperplasia and prostate cáncer

OBJETIVOS: Analizar la presencia delvirus de papiloma humano en próstata y su asociacióncon cáncer.MATERIAL Y MÉTODOS: Se realizó un estudio de casos y controles, para lo cual se colectaronmuestras de tejido con hiperplasia benigna y concáncer de próstata. Se realizó una historia clínicapara conocer la presencia de los factores de riesgoasociados al cáncer de próstata, así como los relacionados con el virus. El tejido prostático fue obtenido por biopsia transrectal o resección transuretral.La identificación del genoma viral se realizó amplificando un fragmento de 450 del gen L1 por mediode una PCR clásica. Para la identificación de los genotipos 16 y 18, se utilizó PCR tiempo real. Para elanálisis de los datos se utilizó la prueba de χ2, prueba de T de Student o U de Mann-Whitney y cálculode OR.RESULTADOS: Se incluyeron 32 pacientes concáncer de próstata y 99 con hiperplasia benigna depróstata en una relación 1:3. La media de edad fuede 69.44±9.22 años. La prevalencia global del virusde papiloma humano fue de 15.2% siendo similar en los casos y entre casos (15.6%) y controles (15.1%),no existiendo diferencia significativa (p=0.572). El40% de las infecciones eran persistentes. Del totalde las muestras positivas, solamente en el 40% seencontró alguno de los dos genotipos analizados (16y 18). Los pacientes con un puntaje de Gleason > 7tuvieron una prevalencia del virus de 16%.CONCLUSIONES: Los resultados ponen de manifiesto la presencia del genoma del virus del papilomahumano en próstata con y sin neoplasia, no se encontróasociación entre la infección y el cáncer de próstata.

OBJECTIVE: To analyze the presenceof human papillomavirus in prostate and its associationwith prostate cancer.METHODS: A case-control study was conducted.Tissue samples with benign hyperplasia and prostatecancer were collected. Risk factors related to prostate cancer and human papillomavirus were assessedby a medical interview. Prostate tissue was obtainedby transrectal biopsy or transurethral resection. Theidentification of viral genome was assessed by the amplification of 450 pb., from L1 gene. Real time PCR wasused to identified HPV genotypes 16 and 18. For dataanalysis, the χ2 test, Student’s T test or Mann-WhitneyU test and OR were computed. RESULTS: Thirty and 99 with benign prostatehyperplasia were included in a 1:3 ratio, with a meanage of 69.44±9.22 years. The global prevalence of human papillomavirus was 15.2% being similar in bothcases (15.6%) and controls (15.1%) with no significantdifference (p = 0.572). Forty percent of the infectionswere persistent. From all positive samples, only in the40% were identified some of the genotypes analyzed(16 and 18). The group of patients with Gleason scorede > 7 had a virus prevalence of 16%CONCLUSIONS: The results show the presence ofthe human papillomavirus genome in prostate tissuewith and without neoplasia; no association was foundbetween infection and prostate cancer.

Human papillomavirus type 13: Genome amplification and characterization data.

As for 2020 only two complete genomes of Human papillomavirus type 13 (HPV13) are publicly available in GenBank database. In addition, reports of partial sequences of genetic regions are very limited. Therefore, genomic research that contributes to knowledge of viral components involved in HPV13 pathogenesis, and molecular mechanisms associated to multifocal epithelial hyperplasia (MEH) disease are urged. In the accompanying paper [1], we aimed to obtain the complete genome sequence of HPV13 associated to MEH disease, obtained from a Mayan boy living in Yucatan, Mexico. Coding sequences were annotated, and viral proteins traduced and deposited in GenBank with accession number MT068446. In this data report, we present the oligonucleotide list used to amplify the complete genome, a graphical abstract of process employed for the amplification of circular HPV13 genome, a representative figure of PCR products obtained for sequencing and multiple sequence alignments with the translated coding sequences of the existing genomes: X62843 is the first HPV13 genome reported [2]; it was generated from a clone obtained from a Turkish patient; DQ344807 was originally obtained from a patient in the Amazonian region [3]. The multiple sequence alignments show the main viral proteins (predicted). This provides relevant information for future molecular analysis and epidemiological studies because HPV13 is an understudied genotype associated to a neglected disease that appears more commonly in children. Additionally, the description of the methods can help in future sequencing of HPV genomes. We hope that our solutions will help researchers who do not have next-generation sequencing (NGS) platforms. A more comprehensive analysis of this data may be obtained from "Genomic characterization of Human papillomavirus type 13, associated to Multifocal Epithelial Hyperplasia, in a Mayan community" [1].

Genomic characterization of human papillomavirus type 13, associated to multifocal epithelial hyperplasia, in a Mayan community.

Human papillomavirus type 13 (HPV13) is a low-risk HPV type associated with Multifocal Epithelial Hyperplasia (MEH). It is considered a rare pathology of oral mucosa, more prevalent in certain ethnical groups, such as the Maya from Yucatan in Mexico. As for 2020 only two complete genomes of HPV13 are publicly available in Genbank database (one from Turkey one from the Amazonian). We aimed to obtain the complete genome sequence of HPV13 associated to MEH, obtained from a community in the Mayan area from Mexico. A bank of oral swabs from children with MEH were used. To enrich the sample, a Rolling Cycle Amplification (RCA) method was performed followed by overlapping end-point PCR of 500 bp fragments, Sanger sequencing and assembly. Eight open reading frames (ORFs) were annotated (E1, E2, E4, E5, E6, E7, L1 and L2 genes). When compared with the other two previously reported genomes the identity at nucleotide level is high 98.9% and 99.6%, respectively. The phylogenetic tree shows that Yucatan HPV13 is more closely related to HPV13 obtained from the Amazonian. Most changes identified at amino acid level are substitutions derived from nucleotide variations or SNPs in coding regions. Amino-acid changes were observed in E2 and E1 proteins (n ≥ 8), and in L1, L2, E6 and E5 proteins (n ≤ 5). E7 protein from Yucatan has 100% identity with the reported from Amazonian and differs (94.1% identity) with the one from Turkey due to 3 substitutions and three missing amino acids. In conclusion, the genome from HPV13 (7831 bp, 49 nt missing) associated to MEH in the Mayan area from Yucatan was obtained from stored swabs; this is the first effort in Mexico, the second in Latin America, and the third of the world. More research that contributes to the knowledge of the determinants underlying this neglected pathology are urged.

Infección por citomegalovirus humano en neonatos de un hospital público de Mérida, Yucatán.

INTRODUCTION: Human cytomegalovirus (HCMV) is recognized as the most common cause of congenital viral infection, which can occur as a result of primary infection, reinfection or infection reactivation in the pregnant woman and be the cause of delay in neuronal development and sensorineural hearing loss in the neonate. OBJECTIVE: To identify CMVH infection in newborns by real-time polymerase chain reaction (RT-PCR) and cell culture. METHOD: Observational, cross-sectional, retrospective study with oral swab samples from 362 neonates born within a 10-month period in a public hospital of Mérida, Yucatán. RT-PCR was carried out for the detection of HCMV. Fibroblast primary cell culture was obtained from human foreskin tissue to isolate the virus. Only positive cases were followed. RESULTS: A prevalence of HCMV infection of 0.86 % was found by RT-PCR. No virus was isolated with cell culture. In the follow-up visits, sensory health and neurodevelopment were adequate. CONCLUSION: The prevalence of HCMV infection is similar to that of worldwide reports, and only was detected by RT-PCR. Asymptomatic infection detected 12-14 h after birth had no long-term health consequences.

INTRODUCCIÓN: El citomegalovirus humano es reconocido como la causa más común de infección viral congénita, la cual puede darse como resultado de infección primaria, reinfección o reactivación en la mujer embarazada; además, puede ocasionar retraso en el desarrollo neuronal y pérdida auditiva sensoneural en el neonato. OBJETIVO: Identificar la infección por citomegalovirus humano en neonatos por PCR en tiempo real (PCR-TR) y cultivo celular. MÉTODO: Estudio observacional, longitudinal y retrospectivo con muestras de hisopado oral provenientes de 362 neonatos nacidos en un periodo de 10 meses en un hospital público de Mérida, Yucatán. Se realizó PCR-TR para la detección de citomegalovirus humano. Se obtuvo cultivo celular primario de fibroblastos a partir de tejido de prepucio humano para recuperar el virus. Se siguieron solo los casos positivos. RESULTADOS: Se encontró 0.86 % de infección por citomegalovirus humano por PCR-TR. No se recuperó el virus en cultivo. En las visitas de seguimiento, la salud sensorial y el neurodesarrollo fueron adecuados. CONCLUSIÓN: La prevalencia de infección por citomegalovirus humano en neonatos fue similar a la de reportes mundiales y solo pudo evidenciarse por PCR. La infección asintomática detectada entre las 12 a 24 horas del nacimiento no tuvo consecuencias a largo plazo.

Infección por citomegalovirus humano en neonatos de un hospital público de Mérida, Yucatán / Human cytomegalovirus infection in neonates from a public hospital in Mérida, Yucatán

Resumen Introducción: El citomegalovirus humano es reconocido como la causa más común de infección viral congénita, la cual puede darse como resultado de infección primaria, reinfección o reactivación en la mujer embarazada; además, puede ocasionar retraso en el desarrollo neuronal y pérdida auditiva sensoneural en el neonato. Objetivo: Identificar la infección por citomegalovirus humano en neonatos por PCR en tiempo real (PCR-TR) y cultivo celular. Método: Estudio observacional, longitudinal y retrospectivo con muestras de hisopado oral provenientes de 362 neonatos nacidos en un periodo de 10 meses en un hospital público de Mérida, Yucatán. Se realizó PCR-TR para la detección de citomegalovirus humano. Se obtuvo cultivo celular primario de fibroblastos a partir de tejido de prepucio humano para recuperar el virus. Se siguieron solo los casos positivos. Resultados: Se encontró 0.86 % de infección por citomegalovirus humano por PCR-TR. No se recuperó el virus en cultivo. En las visitas de seguimiento, la salud sensorial y el neurodesarrollo fueron adecuados. Conclusión: La prevalencia de infección por citomegalovirus humano en neonatos fue similar a la de reportes mundiales y solo pudo evidenciarse por PCR. La infección asintomática detectada entre las 12 a 24 horas del nacimiento no tuvo consecuencias a largo plazo.

Abstract Introduction: Human cytomegalovirus (HCMV) is recognized as the most common cause of congenital viral infection, which can occur as a result of primary infection, reinfection or infection reactivation in the pregnant woman and be the cause of delay in neuronal development and sensorineural hearing loss in the neonate. Objective: To identify CMVH infection in newborns by real-time polymerase chain reaction (RT-PCR) and cell culture. Method: Observational, cross-sectional, retrospective study with oral swab samples from 362 neonates born within a 10-month period in a public hospital of Mérida, Yucatán. RT-PCR was carried out for the detection of HCMV. Fibroblast primary cell culture was obtained from human foreskin tissue to isolate the virus. Only positive cases were followed. Results: A prevalence of HCMV infection of 0.86 % was found by RT-PCR. No virus was isolated with cell culture. In the follow-up visits, sensory health and neurodevelopment were adequate. Conclusion: The prevalence of HCMV infection is similar to that of worldwide reports, and only was detected by RT-PCR. Asymptomatic infection detected 12-14 h after birth had no long-term health consequences.

Prevalence of anal infection due to high-risk human papillomavirus and analysis of E2 gene integrity among women with cervical abnormalities / Prevalencia de infección anal por virus de papiloma humano de alto riesgo y análisis de la integridad del gen E2 en mujeres con anormalidades cervicales

BACKGROUND: High-risk human papillomaviruses (HR-HPV) infection has been associated with 90% of anal cancer cases. Women with abnormal cytology are a high-risk group to develop anal neoplasia. The aim of this study is to describe the prevalence and epidemiology of HR-HPV 16, 18, 45, and 58 anal infections in women with cervical abnormalities, as well as to assess E2 gene integrity. METHODS: A cross-sectional study was performed on 311 cervical and 311 anal samples from patients with abnormal cytology in two colposcopy clinics in Yucatan, Mexico. A specific PCR for oncogenes was performed in order to identify HVP 16, 18, 45 and 58. Real time PCR was used to amplify the whole HPV 16, 18, and 58 E2 gene to verify its integrity in anal samples. RESULTS: High risk HPV 16, 18, 58, and/or 45 were found in 41.47% (129/311) of cervical samples, and in 30.8% (96/331) of anal samples, with 18% (57/311) of the patients being positive in both samples. The same genotypes in both anatomical sites were observed in 11.25% (35/311). The E2 gene was disrupted in 82% of all tested samples. The frequency of genome disruption viral integration in anal samples by genotype was: HPV 58 (97.2%); HPV 16 (72.4%), and HPV 18 (0%). CONCLUSION: Women with cervical disease have HR-HPV anal infections, and most of them have the E2 gene disrupted, which represents a risk to develop anal cancer

ANTECEDENTES: La infección por virus del papiloma humano de alto riesgo (AR-VPH) está asociada al 90% de los casos de cáncer anal; las mujeres con enfermedad cervical son un grupo de alto riesgo para desarrollar neoplasia anal. El objetivo de este estudio es describir la prevalencia y epidemiología de las infecciones anales por AR-VPH 16, 18, 45 y 58 en mujeres con citología anormal y evaluar la integridad del gen E2. MÉTODOS: Se realizó un estudio transversal con 311 muestras cervicales y 311 muestras anales de pacientes con citología anormal de 2 clínicas de colposcopia en Yucatán, México. La identificación de los VPH 16, 18, 45 y 58 se realizó con una PCR específica para los oncogenes. Para verificar la integridad del gen E2 en muestras anales se utilizó PCR en tiempo real para la amplificación de todo el gen E2 de VPH 16, 18 y 58. RESULTADOS: La presencia de AR-VPH 16, 18, 45 y/o 58 fue identificada en el 41,47% (129/311) de las muestras cervicales y en el 30,8% (96/331) de las muestras anales; el 18% de las pacientes (57/311) fueron positivas para ambas muestras, y el 11,25% (35/311) tuvieron el mismo genotipo en ambos sitios anatómicos. El gen E2 se encontró incompleto en el 82% de todas las muestras anales analizadas. La frecuencia de disrupción del genoma viral por genotipos fue: VPH 58 (97, 2%); VPH 16 (72, 4%) y VPH 18 (0%). CONCLUSIÓN: Las mujeres con enfermedad cervical están infectadas con AR-PVH en la región anal y la mayoría presentan disrupción del gen E2, lo que representa un riesgo para desarrollar cáncer anal

Prevalence of anal infection due to high-risk human papillomavirus and analysis of E2 gene integrity among women with cervical abnormalities.

BACKGROUND: High-risk human papillomaviruses (HR-HPV) infection has been associated with 90% of anal cancer cases. Women with abnormal cytology are a high-risk group to develop anal neoplasia. The aim of this study is to describe the prevalence and epidemiology of HR-HPV 16, 18, 45, and 58 anal infections in women with cervical abnormalities, as well as to assess E2 gene integrity. METHODS: A cross-sectional study was performed on 311 cervical and 311 anal samples from patients with abnormal cytology in two colposcopy clinics in Yucatan, Mexico. A specific PCR for oncogenes was performed in order to identify HVP 16, 18, 45 and 58. Real time PCR was used to amplify the whole HPV 16, 18, and 58 E2 gene to verify its integrity in anal samples. RESULTS: High risk HPV 16, 18, 58, and/or 45 were found in 41.47% (129/311) of cervical samples, and in 30.8% (96/331) of anal samples, with 18% (57/311) of the patients being positive in both samples. The same genotypes in both anatomical sites were observed in 11.25% (35/311). The E2 gene was disrupted in 82% of all tested samples. The frequency of genome disruption viral integration in anal samples by genotype was: HPV 58 (97.2%); HPV 16 (72.4%), and HPV 18 (0%). CONCLUSION: Women with cervical disease have HR-HPV anal infections, and most of them have the E2 gene disrupted, which represents a risk to develop anal cancer.

Temporal distribution and genetic variants in influenza A(H1N1)pdm09 virus circulating in Mexico, seasons 2012 and 2013.

The 2012 and 2013 annual influenza epidemics in Mexico were characterized by presenting different seasonal patterns. In 2012 the A(H1N1)pdm09 virus caused a high incidence of influenza infections after a two-year period of low circulation; whereas the 2013 epidemic presented circulation of the A(H1N1)pdm09 virus throughout the year. We have characterized the molecular composition of the Hemagglutinin (HA) and Neuraminidase (NA) genes of the A(H1N1)pdm09 virus from both epidemic seasons, emphasizing the genetic characteristics of viruses isolated from Yucatan in Southern Mexico. The molecular analysis of viruses from the 2012 revealed that all viruses from Mexico were predominantly grouped in clade 7. Strikingly, the molecular characterization of viruses from 2013 revealed that viruses circulating in Yucatan were genetically different to viruses from other regions of Mexico. In fact, we identified the occurrence of two genetic variants containing relevant mutations at both the HA and NA surface antigens. There was a difference on the temporal circulation of each genetic variant, viruses containing the mutations HA-A141T / NA-N341S were detected in May, June and July; whereas viruses containing the mutations HA-S162I / NA-L206S circulated in August and September. We discuss the significance of these novel genetic changes.

Epidemiología de la infección oral por VPH en sujetos jóvenes sanos / Epidemiology of oral HPV infection in young healthy individuals

Resumen Introducción: La infección por VPH es la infección viral de transmisión sexual más frecuente, y se encuentra asociada a diversas neoplasias. Objetivo: Describir la epidemiología, historia natural y factores de riesgo asociados a la infección oral por VPH en adultos jóvenes asintomáticos. Métodos: Se realizó un estudio prospectivo de corte longitudinal, que incluyó sujetos sin patología oral, a los que se les tomó una muestra de la mucosa bucal. A todos los sujetos con resultados positivos se les realizó un nuevo muestreo seis meses después. Se identificó la presencia del virus por RPC; los datos demográficos y de conducta sexual fueron obtenidos con una encuesta que respondieron sin intervención del investigador. Resultados: Se recolectaron 102 muestras de sujetos de 18-26 años de edad, 60 (58,8%) correspondieron al sexo masculino. La prevalencia del virus fue de 6,9%; todos los sujetos positivos tenían vida sexual. Las relaciones sexuales entre personas del mismo sexo fue la única variable asociada a la presencia del virus (p < 0,05). A los seis meses, todos los sujetos habían eliminado al virus. Conclusión: La infección oral por VPH es transitoria y está asociada a relaciones sexuales entre personas del mismo sexo, principalmente mujeres que tienen sexo con mujeres.

Background: HPV infection is the most common sexually transmitted viral infection, and is associated with several neoplasms. Aim: To describe the epidemiology, natural history and risk factors associated with oral HPV infection in asymptomatic young adults. Methods: A prospective and longitudinal study was conducted, including subjects without oral pathology, who were sampled from the oral mucosa. All subjects with positive results were re-sampled 6 months later. The presence of HPV was identified by PCR. Demographic and sexual behavior data were obtained with a survey that was responded without the intervention of the researcher. Results: 102 samples were collected from subject of 18-26 years old, 60 (58.8%) were male. The prevalence of the virus was 6.9%; all positive subjects had active sexual life. Same-gender relationships were the only variable associated with the presence of the virus (p < 0.05). At six months all subjects had eliminated the virus. Conclusion: Oral HPV infection is transient and is associated to same-gender relationships, mainly women who have sex with women.

Influenza seasonality goes south in the Yucatan Peninsula: The case for a different influenza vaccine calendar in this Mexican region.

INTRODUCTION: While vaccination may be relatively straightforward for regions with a well-defined winter season, the situation is quite different for tropical regions. Influenza activity in tropical regions might be out of phase with the dynamics predicted for their hemispheric group thereby impacting the effectiveness of the immunization campaign. OBJECTIVE: To investigate how the climatic diversity of Mexico hinders its existing influenza immunization strategy and to suggest that the hemispheric vaccine recommendations be tailored to the regional level in order to optimize vaccine effectiveness. METHODS: We studied the seasonality of influenza throughoutMexico by modeling virological and mortality data.De-trended time series of each Mexican state were analyzed by Fourier decomposition to describe the amplitude and timing of annual influenza epidemic cycles and to compare with each the timing of the WHO's Northern and Southern Hemispheric vaccination schedule. FINDINGS: The timings of the primary (major) peaks of both virological and mortality data for most Mexican states are well aligned with the Northern Hemisphere winter (December-February) and vaccine schedule. However, influenza peaks in September in the three states of the Yucatan Peninsula. Influenza-related mortality also peaks in September in Quintana Roo and Yucatan whereas it peaks in May in Campeche. As the current timing of vaccination in Mexico is between October and November, more than half of the annual influenza cases have already occurred in the Yucatan Peninsula states by the time the Northern Hemispheric vaccine is delivered and administered. CONCLUSION: The current Northern Hemispheric influenza calendar adopted for Mexico is not optimal for the Yucatan Peninsula states thereby likely reducing the effectiveness of the immunization of the population. We recommend that Mexico tailor its immunization strategy to better reflect its climatologic and epidemiological diversity and adopt the WHO Southern Hemisphere influenza vaccine and schedule for the Yucatan Peninsula.

Human papillomavirus and Chlamydia trachomatis infection in gyneco-obstetric outpatients from a mexican hospital.

CONTEXT: Human papillomavirus (HPV) and Chlamydia trachomatis are the most frequent sexually transmitted infections, usually asymptomatic. Persistent infection with high-risk HPV types and other cofactors such as the concomitant infection with C. trachomatis can represent a higher risk to develop cervical lesions; therefore, screening with sensitive methods could aid to identify women at risk. AIMS: The aim is to determine the prevalence and concurrence of both infections, detected with in-house molecular methods, and to identify the risk factors associated to the infections in Mexican women. SUBJECTS AND METHODS: This was a cross-sectional study including gynaecological-obstetrical medical outpatients from a Social Security Hospital in Southeast Mexico. After informed consent, cervicovaginal samples were collected and tested for HPV and C. trachomatis using polymerase chain reaction (PCR). HPV positives were further tested for high-risk HPV16, 18, 58 and low-risk 11 using real-time PCR. All methods employed were in-house. Data analyses included odds ratios (OR), Chi-square and linear regressions. RESULTS: Women included were 233, aging 15-49 (mean 30 years), 52.8% were pregnant. For HPV and C. trachomatis testing, 230 samples were adequate, resulting in 48 (20.9%) and 15 (6.5%) positives, respectively; 4 (1.7%) were positive to both. The most frequent genotype identified was HPV58 (25% of typified samples). C. trachomatis positives were 73% asymptomatic, none had pelvic inflammatory disease or infertility histories. The only variable associated to HPV infection was the history of previous sexually transmitted disease (OR = 3.69,P= 0.0019). CONCLUSIONS: More than 25% of the population was infected with either agent. We successfully used in-house molecular methodologies for diagnosis and typing, showing HPV and C. trachomatis prevalence consistent to previous reports. Concomitant infections were found, HPV high-risk types were involved in half of these cases, representing a higher risk.

Genetic Variability in E6 and E7 Oncogenes from Human Papillomavirus Type 58 in Mexican Women.

BACKGROUND/AIMS: The study aimed to describe human papillomavirus (HPV) 58 genetic variability in E6 and E7 oncogenes from women in southeast Mexico and their phylogenetic relationships with the sequences from other geographical regions. METHODS: The E6-E7 region was amplified by nested PCR, and sequenced for identification of polymorphisms, phylogenetic trees construction, and haplotype and fixation tests. RESULTS: HPV58 positive samples were obtained from a repository, 54 were amplified, 47 sequences for the E6 gene, and 51 sequences for the E7 gene were obtained. Fifteen new E6 mutations were found; the most frequent were G279T (G57V; 29.78%), T249G (F47C; 34.04%), and A270G (Y54C; 34.04%), and previously reported c307t (63.82%). For E7, 17 known mutations were found, the most frequent were C632T (T20I), 35.30%, G760A (G63S), 35.30%, and t744g 74.50%. No significant association with the severity of the lesions was found. The polytomy in the E6 tree did not allow proposing phylogenetic relationship, and E7 tree presented defined branches. All sequences were presumably A lineage, most closely related to A1 and/or A3 sublineage. HPV58 variants are not specific for a geographical area. Population and fixation analyses suggest a possible Asian origin of HPV58 from Yucatan. The most frequent E7 haplotype in Yucatan groups with other populations of the world. CONCLUSION: The genetic variability of HPV58 from Mexico was described for the first time. E7 was more conserved than E6. New mutants present exclusively in Yucatan were identified.

[Epidemiology of oral HPV infection in young healthy individuals]. / Epidemiología de la infección oral por VPH en sujetos jóvenes sanos.

BACKGROUND: HPV infection is the most common sexually transmitted viral infection, and is associated with several neoplasms. AIM: To describe the epidemiology, natural history and risk factors associated with oral HPV infection in asymptomatic young adults. METHODS: A prospective and longitudinal study was conducted, including subjects without oral pathology, who were sampled from the oral mucosa. All subjects with positive results were re-sampled 6 months later. The presence of HPV was identified by PCR. Demographic and sexual behavior data were obtained with a survey that was responded without the intervention of the researcher. RESULTS: 102 samples were collected from subject of 18-26 years old, 60 (58.8%) were male. The prevalence of the virus was 6.9%; all positive subjects had active sexual life. Same-gender relationships were the only variable associated with the presence of the virus (p < 0.05). At six months all subjects had eliminated the virus. CONCLUSION: Oral HPV infection is transient and is associated to same-gender relationships, mainly women who have sex with women.

Analysis of E2 gene integrity in HPV16 and HPV58 viruses isolated from women with cervical pathology.

Integration of human papillomavirus (HPV) DNA into human cells accompanied by the disruption of the viral genome has been described as a prerequisite for cancer development. This study aimed to investigate E2 gene integrity of HPV16 and HPV58 viruses isolated from infected women with cervical lesions. Forty-two HPV16- and 31 HPV58-positive samples were analysed. E2 integrity was assumed when all fragments covering the E2 gene were amplified with specific polymerase chain reaction primers. Overall, in 59% of the samples, at least one fragment was not amplified in HPV16- (57%) and HPV58-positive samples (61%). Samples from high-grade squamous intraepithelial lesions had the highest frequency of E2 gene disruptions (73%), followed by samples from low-grade squamous intraepithelial lesions (63%) and, finally, samples from invasive cervical cancer (35%). Association between the integrity status of the E2 gene, and lesion grade was assessed by the chi-squared test applied to the combined set of viruses (p = 0.6555) or to populations of the same virus type (HPV58, p = 0.3101; HPV16, p = 0.3024). In conclusion, in this study, no association was found between the presence of E2 gene disruptions and the grade of cervical lesions caused by HPV16 and HPV58.

Mutations at highly conserved residues in influenza A(H1N1)pdm09 virus affect neuraminidase activity.

Influenza virus neuraminidase (NA) plays a pivotal role during viral growth since its sialidase activity allows the efficient release of nascent virions from infected cells. Consequently, mutations in the NA catalytic site affecting sialic acid (SA) cleavage may influence the biological properties of influenza viruses. This study reports two amino acid substitutions (N386K and P431S) in the NA of the influenza A(H1N1)pdm09 virus that emerged in 2009 in Mexico. The NA sialidase activity to cleave SA-like substrates, and viral growth were examined and the mutant viruses had various deficiencies. NA mutations N386K and P431S together or separately, and in the presence or absence of H275Y were further evaluated using recombinant influenza A/California/04/2009 (pH1N1) viruses containing single, double, or triple mutations. Viral growth was reduced in the presence of mutation P431S alone or combined with N386K and/or H275Y. Substrates hydrolysis was reduced when recombinant pH1N1 viruses were analyzed by NA inhibitory assays. Moreover, elution assays with guinea pig red blood cells indicated an unbalanced hemagglutinin (HA):NA functionality. Altogether, our data underline the functional significance of mutations at highly conserved sites in influenza virus NA glycoprotein and the occurrence of permissive mutations to compensate virus viability in vitro.

Epidemiology of oral HPV in the oral mucosa in women without signs of oral disease from Yucatan, Mexico.

High-risk human papillomaviruses (HR-HPV) are considered necessary for the development of cervical cancer. Furthermore, there is no doubt that some types of oral squamous cell carcinoma are associated with HR-HPV. The epidemiology of oral HPV infections in healthy subjects remains unclear due to a lack of knowledge. The objective of this study was to investigate the epidemiology of human papillomavirus infections of the oral mucosa without pathology. A cross-sectional study was performed; samples from 390 women seeking prenatal care, Pap smears, family planning or gynecological diseases were studied. Oral cells were collected by direct swab sampling. Information regarding sociodemographic status, sexual behavior, infectious diseases, contraceptive history and tobacco and alcohol consumption were obtained through direct interviews. HPV and genotypes were detected by type-specific polymerase chain reaction. Our results revealed that 14% of the women studied had an oral HPV infection. Women ≤ 20 years of age had the highest HPV prevalence (24.5%). In total, seven genotypes were identified, including the high-risk genotypes 16, 18, 58 and 59 and the low-risk genotypes 6, 81 and 13, the latter of which is a type exclusive to oral mucosa. Sexual behavior was not associated with the presence of genital HPV types in the oral mucosa. Genital HPV types were present in the oral mucosa of women without associated clinical manifestations; however, sexual behavior was not associated with infection, and therefore others routes of transmission should be explored.