First report of natural Wolbachia infections in mosquitoes from Cuba.

Mosquitoes are extensively responsible for the transmission of pathogens. Novel strategies using Wolbachia could transform that scenario, since these bacteria manipulate mosquito reproduction, and can confer a pathogen transmission-blocking phenotype in culicids. Here, we screened the Wolbachia surface protein region by PCR in eight Cuban mosquito species. We confirmed the natural infections by sequencing and assessed the phylogenetic relationships among the Wolbachia strains detected. We identified four Wolbachia hosts: Aedes albopictus, Culex quinquefasciatus, Mansonia titillans, and Aedes mediovittatus (first report worldwide). Knowledge of Wolbachia strains and their natural hosts is essential for future operationalization of this vector control strategy in Cuba.

Emergence and evolution of SARS-CoV-2 genetic variants during the Cuban epidemic.

The SARS CoV-2 D614G variant circulated in Cuba in 2020. New viral variants were detected after the opening of the border in November 2020. We show the results of the genomic surveillance in Cuba from December 28, 2020, to September 28, 2021 and their relationship to the epidemiological situation in the country. A total of 1,406 nasopharyngeal exudates from COVID-19 patients were processed for RNA extraction and the 1836 bp fragment of the spike gene was amplified and sequenced. The mutations present were determined using the GISAID database. Prevalence ratios were estimated by fitting Poisson univariate and multivariate regression models to investigate associations between SARS-CoV-2 variant group (VOC, non-VOC) and disease outcome. Seventeen genetic variants were detected including VOC Alpha, Beta, Gamma and Delta, one variant of interest (VOI) (Lambda) and two previous VOI (A.2.5.1 and Zeta/P.2). Beta (34.77%), Delta (24.89%) and D614G (19%) variants were the most frequently detected. By June, Delta increased in frequency, displacing Beta. Disease severity increased significantly with age and VOC (PR =1.98, IC 95%: 1.33-3.05, p <0.05). Genomic surveillance allowed us to identify the upsurge of novel variants. Coinciding with the higher epidemic period, multiple variants were co-circulating. Although we cannot rule out that failure in the transmission containment measures occurred, the increase in the number of cases associated with the circulation of several variants, particularly the Beta and Delta variants is highly suggestive. A greater association of Beta variant with clinical severity and Delta variant with a greater transmissibility was observed.

Correction: Cuban history of CRF19 recombinant subtype of HIV-1.

[This corrects the article DOI: 10.1371/journal.ppat.1009786.].

Cuban history of CRF19 recombinant subtype of HIV-1.

CRF19 is a recombinant form of HIV-1 subtypes D, A1 and G, which was first sampled in Cuba in 1999, but was already present there in 1980s. CRF19 was reported almost uniquely in Cuba, where it accounts for ∼25% of new HIV-positive patients and causes rapid progression to AIDS (∼3 years). We analyzed a large data set comprising ∼350 pol and env sequences sampled in Cuba over the last 15 years and ∼350 from Los Alamos database. This data set contained both CRF19 (∼315), and A1, D and G sequences. We performed and combined analyses for the three A1, G and D regions, using fast maximum likelihood approaches, including: (1) phylogeny reconstruction, (2) spatio-temporal analysis of the virus spread, and ancestral character reconstruction for (3) transmission mode and (4) drug resistance mutations (DRMs). We verified these results with a Bayesian approach. This allowed us to acquire new insights on the CRF19 origin and transmission patterns. We showed that CRF19 recombined between 1966 and 1977, most likely in Cuban community stationed in Congo region. We further investigated CRF19 spread on the Cuban province level, and discovered that the epidemic started in 1970s, most probably in Villa Clara, that it was at first carried by heterosexual transmissions, and then quickly spread in the 1980s within the "men having sex with men" (MSM) community, with multiple transmissions back to heterosexuals. The analysis of the transmission patterns of common DRMs found very few resistance transmission clusters. Our results show a very early introduction of CRF19 in Cuba, which could explain its local epidemiological success. Ignited by a major founder event, the epidemic then followed a similar pattern as other subtypes and CRFs in Cuba. The reason for the short time to AIDS remains to be understood and requires specific surveillance, in Cuba and elsewhere.

Recomendaciones para el abordaje del paro cardiorrespiratorio en niños durante la pandemia de la COVID-19 / Recommendations to tackle cardiorespiratory arrest in children during COVID-19 pandemic

La parada cardiorrespiratoria no es un evento frecuente en pediatría, pero para aquellos que la sufren, representa un serio problema por la elevada mortalidad o el riesgo de secuelas. La causa más frecuente es la hipoxia, por tanto, las intervenciones respiratorias son imprescindibles durante la reanimación cardiopulmonar. Debido a la pandemia de la COVID-19, el abordaje de la vía aérea durante la resucitación ha sido analizada teniendo en cuenta el alto riesgo de contagiosidad del virus SARS-CoV-2 por la aerosolización de las secreciones respiratorias. Se recomienda el uso de equipos de protección, limitar el personal necesario para la resucitación, preferir la ventilación por tubo endotraqueal si hay personal entrenado y extremar la vigilancia del paciente pediátrico en riesgo para compensar la demora por la colocación de los medios de protección. Es imprescindible conocer cada una de las intervenciones en los distintos eslabones de la cadena de supervivencia para lograr la recuperación del paciente con el menor número de secuelas posibles y sin contagio en el personal de la salud(AU)

Cardiorespiratory arrest is a not a frequent event in pediatrics, but it is a serious problem for those who suffer it due to its high mortality rates and the risk of sequelaes. Hypoxia is its most frequent cause that is why respiratory interventions are essential during cardiopulmonary resuscitation. Due to COVID-19 pandemic, tackling the air ways during resuscitation has been subject of analysis taking into account the high risk of contagion of the virus SARS-COV-2 by the aerosolization of respiratory secretions. It is recommended the use of protection equipment, to limit personnel to the just needed for resuscitation, to choose ventilation by endotracheal intubation if there is trained personnel, and to maximize the surveillance of paediatric patients at risk in order to compensate the delay due to the placing of protection means. It is essential to know each of the interventions in the different links of the survival chain in order to achieve the patient's recovery with the lower number of possible sequelaes and without contagions among the health personnel(AU)

Presentación de una paciente con embarazo ectópico ovárico / Presentation of a Patient with Ovarian Ectopic Pregnancy

El embarazo ectópico es en la actualidad un problema de salud a nivel mundial, atribuido el aumento de su incidencia, a las inflamaciones pélvicas provocadas por enfermedades de transmisión sexual, el aborto, uso de dispositivos intrauterinos, la aplicación de técnicas de reproducción asistida, uso de inductores de la ovulación, entre otros. Se presentó un caso de embarazo ectópico ovárico en una mujer de la raza negra de 38 años de edad, que acudió con cuadro clínico de vómitos, escalofríos y dolor abdominal al Cuerpo de Guardia del Margret Marquart Catholic Hospital en Kpando, región Volta, Ghana. Al examen físico se palpó una tumoración que ocupaba gran parte del hipogastrio, fondo de saco de Douglas abombado y doloroso. Con antecedentes patológicos personales de anemia crónica y portadora de virus de inmunodeficiencia humana (VIH). Se ingresó, se realizó laparotomía y se egresó luego de evolución satisfactoria.

Ectopic pregnancy is nowadays a worldwide health problem due to the incidence of pelvic inflammatory diseases caused by abortion, sexually transmitted diseases, the use of intrauterine devices and assisted reproductive techniques among others. A 38- year- old black woman came to the emergency room at the Margret Marquart Catholic Hospital of Kpando, Volta Region in Ghana with a history of vomiting, chills and abdominal pain. On physical examination, a tumor mass was found, that occupied a large part of the epigastrium, Douglas sack fundus, which was convex and painful. She has a personal pathological history of chronic anaemia and she was a carrier of HIV. She was admitted in the hospital and a laparotomy was performed. She was discharged after her satisfactory evolution.

Presentación de una paciente con embarazo ectópico ovárico / Presentation of a Patient with Ovarian Ectopic Pregnancy

El embarazo ectópico es en la actualidad un problema de salud a nivel mundial, atribuido el aumento de su incidencia, a las inflamaciones pélvicas provocadas por enfermedades de transmisión sexual, el aborto, uso de dispositivos intrauterinos, la aplicación de técnicas de reproducción asistida, uso de inductores de la ovulación, entre otros. Se presentó un caso de embarazo ectópico ovárico en una mujer de la raza negra de 38 años de edad, que acudió con cuadro clínico de vómitos, escalofríos y dolor abdominal al Cuerpo de Guardia del Margret Marquart Catholic Hospital en Kpando, región Volta, Ghana. Al examen físico se palpó una tumoración que ocupaba gran parte del hipogastrio, fondo de saco de Douglas abombado y doloroso. Con antecedentes patológicos personales de anemia crónica y portadora de virus de inmunodeficiencia humana (VIH). Se ingresó, se realizó laparotomía y se egresó luego de evolución satisfactoria(AU)

Ectopic pregnancy is nowadays a worldwide health problem due to the incidence of pelvic inflammatory diseases caused by abortion, sexually transmitted diseases, the use of intrauterine devices and assisted reproductive techniques among others. A 38- year- old black woman came to the emergency room at the Margret Marquart Catholic Hospital of Kpando, Volta Region in Ghana with a history of vomiting, chills and abdominal pain. On physical examination, a tumor mass was found, that occupied a large part of the epigastrium, Douglas sack fundus, which was convex and painful. She has a personal pathological history of chronic anaemia and she was a carrier of HIV. She was admitted in the hospital and a laparotomy was performed. She was discharged after her satisfactory evolution(AU)

Human papillomavirus infection in anal intraepithelial lesions from HIV infected Cuban men.

BACKGROUND: An association between HPV infection and progression to anal squamous intraepithelial lesions (ASIL) has been established, specifically in high-risk populations such as HIV-infected men. In this population, anal cancer is one of the most common non-AIDS-defining malignancies. METHODS: A cross-sectional study to detect anal lesions and HPV infection was performed. Anal mucosa samples were collected from 56 HIV-infected men from Cuba. The cytological diagnosis was done according to Bethesda 2001 System. HPV DNA detection was determined by qPCR for six high-risk HPV types and end point PCR for low-risk HPV types (6 and 11). The end point PCR with nucleotide sequencing technique was achieved to detect other genotypes of HPV not included in the qPCR in those samples negative for HPV- 6 and 11 or negative for the six genotypes identified in the qPCR. RESULTS: Cytological diagnosis identified 53 of 56 (95%) men with abnormal anal cytology. Among those, 26% (14/53) had atypical squamous cells of undetermined significance (ASC-US), 4% (2/53) had atypical squamous cells of undetermined significance cannot exclude high-grade lesions (ASC-H), 64% (34/53) had low-grade squamous intraepithelial lesions (LSIL), and 6% (3/53) had high-grade squamous intraepithelial lesions (HSIL). HPV DNA was detected in 89% (50/56) of men and 79% had at least one of the high-risk HPV types. HPV- 16 was the most common genotype (52%), while HPV-18 was the most frequently detected genotype in men with HSIL. We found statistically significant differences in the HPV viral loads with respect to the cytology results (p = 0.0006) and that the practice of receptive anal sex was a risk factor for anal HPV infection (p = 0.032). CONCLUSION: This study shows a high prevalence of ASIL and high-risk HPV infections in the study group and is the first study showing the distribution of HPV genotypes in HIV infected Cuban men with abnormal anal cytology. This information may be of importance for local decision makers to improve prevention strategies, including the introduction of HPV vaccine in Cuba.

Etiology of Genital Ulcer Disease in Male Patients Attending a Sexually Transmitted Diseases Clinic: First Assessment in Cuba.

BACKGROUND: Sexually transmitted diseases (STDs) and in particular genital ulcer disease (GUD) have a major impact on morbidity and mortality in developing countries. The World Health Organization recommends the use of syndromic guidelines for the treatment of sexually transmitted infections (STIs) in resource-constrained countries. Surveillance of autochthonous etiologies provides epidemiological information contributing to the prevention and treatment of STIs. We investigated the etiology and factors associated with GUD among male patients attending a STD clinic in Havana, Cuba. METHODS: Swabs from genital ulcers of 113 male patients, collected from May 2012 to June 2015, were analyzed using PCR for herpes simplex virus types 1 and 2, Treponema pallidum, Haemophilus ducreyi, and Chlamydia trachomatis. We also investigated the clinical and epidemiological characteristics associated with the presence of these pathogens in GUD. RESULTS: At least one of the pathogens was detected in 70% of patients. The occurrence of the pathogens was herpes simplex virus type 2 (HSV-2) (51.3%), T. pallidum (29.2%), and C. trachomatis (1.8%). Co-infections occurred as follows: T. pallidum-HSV-2 (10.6%), C. trachomatis-HSV-2 (0.9%) and C. trachomatis-T. pallidum (0.9%). Herpes simplex virus type 1 and H. ducreyi were not detected. Ages 15 to 40 years, HIV-positive serostatus, and no condom use were significant risk factors for the presence of HSV-2 in genital ulcers. CONCLUSIONS: Our preliminary results highlight the predominance of HSV-2 and T. pallidum as the leading GUD etiologies in the study population and identified risk factors associated with HSV-2. This information should help to inform guidelines for better management of GUD in Havana, Cuba.

CRF19_cpx is an Evolutionary fit HIV-1 Variant Strongly Associated With Rapid Progression to AIDS in Cuba.

BACKGROUND: Clinicians reported an increasing trend of rapid progression (RP) (AIDS within 3 years of infection) in Cuba. METHODS: Recently infected patients were prospectively sampled, 52 RP at AIDS diagnosis (AIDS-RP) and 21 without AIDS in the same time frame (non-AIDS). 22 patients were sampled at AIDS diagnosis (chronic-AIDS) retrospectively assessed as > 3 years infected. Clinical, demographic, virological, epidemiological and immunological data were collected. Pol and env sequences were used for subtyping, transmission cluster analysis, and prediction of resistance, co-receptor use and evolutionary fitness. Host, immunological and viral predictors of RP were explored through data mining. FINDINGS: Subtyping revealed 26 subtype B strains, 6 C, 6 CRF18_cpx, 9 CRF19_cpx, 29 BG-recombinants and other subtypes/URFs. All patients infected with CRF19 belonged to the AIDS-RP group. Data mining identified CRF19, oral candidiasis and RANTES levels as the strongest predictors of AIDS-RP. CRF19 was more frequently predicted to use the CXCR4 co-receptor, had higher fitness scores in the protease region, and patients had higher viral load at diagnosis. INTERPRETATION: CRF19 is a recombinant of subtype D (C-part of Gag, PR, RT and nef), subtype A (N-part of Gag, Integrase, Env) and subtype G (Vif, Vpr, Vpu and C-part of Env). Since subtypes D and A have been associated with respectively faster and slower disease progression, our findings might indicate a fit PR driving high viral load, which in combination with co-infections may boost RANTES levels and thus CXCR4 use, potentially explaining the fast progression. We propose that CRF19 is evolutionary very fit and causing rapid progression to AIDS in many newly infected patients in Cuba.

Presentación de una paciente con embarazo heterotópico / Presentation of a Patient with Heterotopic Pregnancy

En los últimos años el embarazo heterotópico ha aumentado, y guarda relación con el desarrollo que alcanzan los tratamientos con técnicas de fertilización in vitro. Aún cuando su etiología no es plenamente esclarecida, muchos de los factores son compartidos con los autores que refieren el embarazo ectópico, como la enfermedad pélvica inflamatoria, factor etiológico potencial en aproximadamente el 45 % de los casos de embarazo ectópico. En el siglo XXI estos tipos de casos son exóticos, sin factores de riesgo aparentes. Se presentó un caso de embarazo heterotópico espontáneo en una mujer de la raza negra de 35 años de edad con antecedentes de transfusión sanguínea por anemia aguda y paridad satisfactoria, que acudió al Cuerpo de Guardia del Margret Marquart Catholic, Hospital en Kpando, región Volta, Ghana; por presentar dolor abdominal, más intenso en hipogastrio. Se realizó laparotomía y se egresó luego de evolución satisfactoria.

In recent years the heterotopic pregnancy has increased, and is related to the development reaching treatments in vitro fertilization techniques. Although its etiology is not fully elucidated, many of the factors are shared with the authors refer ectopic pregnancy, and pelvic inflammatory disease potential in approximately 45% of cases of ectopic pregnancy etiologic factor. In the XXI century, these types of cases are exotic, without apparent risk factors. A case of spontaneous heterotopic pregnancy was presented in a 35-year-old black woman with a history of blood transfusion for severe anemia and satisfactory parity, attended at Emergency Service of Margret Marquart Catholic Hospital in Kpando, Volta region, Ghana was presented in this paper. The patient had severe abdominal pain in lower abdomen. Laparotomy was performed and the patient was discharged after satisfactory outcome.

Presentación de una paciente con embarazo heterotópico / Presentation of a Patient with Heterotopic Pregnancy

En los últimos años el embarazo heterotópico ha aumentado, y guarda relación con el desarrollo que alcanzan los tratamientos con técnicas de fertilización in vitro. Aún cuando su etiología no es plenamente esclarecida, muchos de los factores son compartidos con los autores que refieren el embarazo ectópico, como la enfermedad pélvica inflamatoria, factor etiológico potencial en aproximadamente el 45 % de los casos de embarazo ectópico. En el siglo XXI estos tipos de casos son exóticos, sin factores de riesgo aparentes. Se presentó un caso de embarazo heterotópico espontáneo en una mujer de la raza negra de 35 años de edad con antecedentes de transfusión sanguínea por anemia aguda y paridad satisfactoria, que acudió al Cuerpo de Guardia del Margret Marquart Catholic, Hospital en Kpando, región Volta, Ghana; por presentar dolor abdominal, más intenso en hipogastrio. Se realizó laparotomía y se egresó luego de evolución satisfactoria.(AU)

In recent years the heterotopic pregnancy has increased, and is related to the development reaching treatments in vitro fertilization techniques. Although its etiology is not fully elucidated, many of the factors are shared with the authors refer ectopic pregnancy, and pelvic inflammatory disease potential in approximately 45% of cases of ectopic pregnancy etiologic factor. In the XXI century, these types of cases are exotic, without apparent risk factors. A case of spontaneous heterotopic pregnancy was presented in a 35-year-old black woman with a history of blood transfusion for severe anemia and satisfactory parity, attended at Emergency Service of Margret Marquart Catholic Hospital in Kpando, Volta region, Ghana was presented in this paper. The patient had severe abdominal pain in lower abdomen. Laparotomy was performed and the patient was discharged after satisfactory outcome.(AU)

Performance of an in-house human immunodeficiency virus type 1 genotyping system for assessment of drug resistance in Cuba.

As commercial human immunodeficiency virus type 1 drug resistance assays are expensive, they are not commonly used in resource-limited settings. Hence, a more affordable in-house procedure was set up taking into account the specific epidemiological and economic circumstances of Cuba. The performance characteristics of the in-house assay were evaluated using clinical samples with various subtypes and resistance patterns. The lower limit of amplification was determined on dilutions series of 20 clinical isolates and ranged from 84 to 529 RNA copies/mL. For the assessment of trueness, 14 clinical samples were analyzed and the ViroSeq HIV-1 Genotyping System v2.0 was used as the reference standard. The mean nucleotide sequence identity between the two assays was 98.7% ± 1.0. Additionally, 99.0% of the amino acids at drug resistance positions were identical. The sensitivity and specificity in detecting drug resistance mutations was respectively 94.1% and 99.5%. Only few discordances in drug resistance interpretation patterns were observed. The repeatability and reproducibility were evaluated using 10 clinical samples with 3 replicates per sample. The in-house test was very precise as nucleotide sequence identity among paired nucleotide sequences ranged from 98.7% to 99.9%. The acceptance criteria were met by the in-house test for all performance characteristics, demonstrating a high degree of accuracy. Subsequently, the applicability in routine clinical practice was evaluated on 380 plasma samples. The amplification success rate was 91% and good quality consensus sequences encoding the entire protease and the first 335 codons in reverse transcriptase could be obtained for 99% of the successful amplicons. The reagent cost per sample using the in-house procedure was around € 80 per genotyping attempt. Overall, the in-house assay provided good results, was feasible with equipment and reagents available in Cuba and was half as expensive as commercial assays.

High frequency of antiviral drug resistance and non-b subtypes in HIV-1 patients failing antiviral therapy in Cuba.

INTRODUCTION: Emergence of HIV-1 drug resistance may limit the sustained benefits of antiretroviral therapy (ART) in settings with limited laboratory monitoring and drug options. The objective is to implement the surveillance of drug resistance and subtypes in HIV-1 patients failing ART in Cuba. METHODS: This study compiled clinical and genotypic drug resistance data 588 ART-experienced HIV-1 patients attending a clinical center in Havana in 2009-2013. Drug resistance testing was performed as part of routine clinical care. Drug resistance mutations and levels were determined using Rega version 8.0.2. RESULTS: Eighty-three percent received solely ART containing at least three drugs. Patients from 2009 to 2010 were longer treated (median: 4.9 vs 2.7 years) and exposed to more ART regimens (median: 4 vs 2 regimens) compared to patients from 2011-2013. Nucleoside reverse transcriptase inhibitor (NRTI), non-nucleoside RTI (NNRTI) and PI mutations were present in 83.5, 77.4 and 52.0%. Full-class resistance (FCR) to NRTI, NNRTI, PI and multidrug resistance (MDR) were detected in 25.0, 33.7, 11.4 and 6.3%. FCR to NRTI, NNRTI, PI and MDR were present in 12.8, 28.7, 0 and 0% after first-line failure (164 patients) and in 23.1, 34.6, 3.8 and 3.1% after second-line failure (130 patients). Subtype B (32.5%), BG recombinants (19.6%) and CRF19_cpx (16.2%) were the most prevalent genetic forms. Subtype distribution did not change significantly between 2009-2010 and 2011-2013, except for BG recombinants that increased from 12.2 to 21.3% (p=0.002). CONCLUSIONS: Our study found a high prevalence of drug resistance and supports the need for appropriate laboratory monitoring in clinical practice and access to drug options in case of virological failure.

Presentación atípica de una metástasis ósea del pulmón / Unusual occurrence of osseous metastasis of the lung

Se presenta un paciente masculino de 49 años con antecedentes de Neoplasia de pulmón el cual acude por dolor y aumento de volumen en el miembro inferior izquierdo. Los estudios imaginológicos (radiografías, tomografía computarizada, gammagrafía) sugerían una miositis osificante del tercio medio de la diáfisis femoral izquierda, debido al compromiso de partes blandas, ya que es poco frecuente visualizarlo como una metástasis, pero el diagnóstico histopatológico fue el de una lesión metastásica(AU)

Here is the case of a 49 years-old male patient with a history of lung neoplasia that went to the doctor's because of pain and inflammation of his left leg. Imaging studies including X-rays, CT and scintigraphy indicated ossifying myositis in the medial third of the left femoral diaphysis due to compromised soft tissues, but the histopathological diagnosis showed a metastatic injure(AU)

Presentación atípica de una metástasis ósea del pulmón / Présentation atypique d'une métastase osseuse de poumon / Unusual occurrence of osseous metastasis of the lung

Se presenta un paciente masculino de 49 años con antecedentes de Neoplasia de pulmón el cual acude por dolor y aumento de volumen en el miembro inferior izquierdo. Los estudios imaginológicos (radiografías, tomografía computarizada, gammagrafía) sugerían una miositis osificante del tercio medio de la diáfisis femoral izquierda, debido al compromiso de partes blandas, ya que es poco frecuente visualizarlo como una metástasis, pero el diagnóstico histopatológico fue el de una lesión metastásica(AU)

Here is the case of a 49 years-old male patient with a history of lung neoplasia that went to the doctor's because of pain and inflammation of his left leg. Imaging studies including X-rays, CT and scintigraphy indicated ossifying myositis in the medial third of the left femoral diaphysis due to compromised soft tissues, but the histopathological diagnosis showed a metastatic injure(AU)

Un patient âgé de 49 ans, avec des antécédents de néoplasie de poumon, est vu en consultation due à une douleur et à un grossissement du membre inférieur gauche. L'imagerie (radiographie, tomographie axiale informatisée, scintigraphie) a suggéré une myosite ossifiante au niveau du tiers moyen de la diaphyse fémorale gauche due à une lésion des parties molles. Puisque la métastase est difficile à distinguer, un test histologique a confirmé la lésion métastatique(AU)

Prospective, comprehensive, and effective viral monitoring in Cuban children undergoing solid organ transplantation.

PURPOSE: In Cuba, viral monitoring in the post-transplant period was not routinely performed. The aim of this research is to identify the most frequent viruses that affect transplanted Cuban children, by implementing a viral follow-up during the post-transplant period. METHODS: The study population included all Cuban pediatric patients who underwent solid organ transplantation (SOT) between November 2009 and December 2012. A total of 34 transplanted pediatric patients of kidney (n = 11) and liver (n = 23) were prospectively monitored during a 34-week period for viral DNAemia and DNAuria by simultaneous detection of cytomegalovirus (CMV), Epstein-Barr virus, herpes simplex virus type 1 and 2, varicella zoster virus, human herpesvirus 6, human adenovirus, and polyomaviruses (BKV and JCV) using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Viral genome of at least one virus was detected in 21 of 34 recipients, 18 patients excreted virus in urine while 12 presented DNAemia. CMV (41.2%) and BKV (35.3%) were the most frequent viruses detected during the follow-up. CMV was the virus mainly associated with clinical symptoms and DNAemia. Its excretion in urine (with cut off value of 219 copies/mL) was associated with detection in plasma (p < 0.001); furthermore, CMV viruria was predictive of CMV viremia (OR:8.4, CI:2.4-29.1, p = 0.001). There was no association between high viral load and clinical complications, due to the prompt initiation of preemptive ganciclovir. CONCLUSION: This comprehensive viral monitoring program effectively prevents the development of critical viral disease, thus urge the implementation of qRT-PCR as routine for viral monitoring of transplanted Cuban organ recipients.

Molecular epidemiology of human papillomavirus infections in cervical samples from cuban women older than 30 years.

OBJECTIVE: This study aimed to provide information about the molecular epidemiology of human papillomavirus (HPV) in a group of Cuban women. MATERIALS AND METHODS: DNA from cervical samples was analyzed using a quantitative real-time polymerase chain reaction (PCR), which detects 6 of the clinically most relevant high-risk HPV types. Furthermore, end point PCR and sequencing were performed. Three hundred twenty-two women (211 with positive and 111 with negative cytologic results) aged between 30 and 69 years were enrolled. Risk factors associated with HPV infections and premalignant lesions were also investigated. RESULTS: HPV DNA was detected in 76.1% (245/322) of the studied population, and 34 different genotypes were found. There was an association between HPV infection and low educational level, history of oral contraceptives, menopausal stage, as well as cigarette and/or alcohol consumption. Besides, in a multivariate analysis, previous positive Pap test result and positive colposcopy finding were both predictor variables for HPV infections and for premalignant lesions. Human papillomavirus infection was found in 94.3% of women (199/211) with positive cytologic result and in 41.4% (46/111) of those with negative results, being more likely that the first group was infected with any HPV (odds ratio = 23.43; 95% CI = 11.70-46.92; p = .000). The most common genotypes were HPV types 16, 18, 31, 58, 33, and 45. All the cases with HPV positive findings had at least 1 high-risk HPV genotype. CONCLUSIONS: This is the first report of the molecular epidemiology of HPV in Cuban women, based on results from a DNA sequence and quantitative PCR. Most individuals were infected with high-risk HPV types. These findings support the inclusion of HPV vaccine in Cuba.

Antiretroviral drug resistance in HIV-1 therapy-naive patients in Cuba.

In Cuba, antiretroviral therapy rollout started in 2001 and antiretroviral therapy coverage has reached almost 40% since then. The objectives of this study were therefore to analyze subtype distribution, and level and patterns of drug resistance in therapy-naive HIV-1 patients. Four hundred and one plasma samples were collected from HIV-1 therapy-naive patients in 2003 and in 2007-2011. HIV-1 drug resistance genotyping was performed in the pol gene and drug resistance was interpreted according to the WHO surveillance drug-resistance mutations list, version 2009. Potential impact on first-line therapy response was estimated using genotypic drug resistance interpretation systems HIVdb version 6.2.0 and Rega version 8.0.2. Phylogenetic analysis was performed using Neighbor-Joining. The majority of patients were male (84.5%), men who have sex with men (78.1%) and from Havana City (73.6%). Subtype B was the most prevalent subtype (39.3%), followed by CRF20-23-24_BG (19.5%), CRF19_cpx (18.0%) and CRF18_cpx (10.3%). Overall, 29 patients (7.2%) had evidence of drug resistance, with 4.0% (CI 1.6%-4.8%) in 2003 versus 12.5% (CI 7.2%-14.5%) in 2007-2011. A significant increase in drug resistance was observed in recently HIV-1 diagnosed patients, i.e. 14.8% (CI 8.0%-17.0%) in 2007-2011 versus 3.8% (CI 0.9%-4.7%) in 2003 (OR 3.9, CI 1.5-17.0, p=0.02). The majority of drug resistance was restricted to a single drug class (75.8%), with 55.2% patients displaying nucleoside reverse transcriptase inhibitor (NRTI), 10.3% non-NRTI (NNRTI) and 10.3% protease inhibitor (PI) resistance mutations. Respectively, 20.7% and 3.4% patients carried viruses containing drug resistance mutations against NRTI+NNRTI and NRTI+NNRTI+PI. The first cases of resistance towards other drug classes than NRTI were only detected from 2008 onwards. The most frequent resistance mutations were T215Y/rev (44.8%), M41L (31.0%), M184V (17.2%) and K103N (13.8%). The median genotypic susceptibility score for the commonly prescribed first-line therapies was 2.5. This analysis emphasizes the need to perform additional surveillance studies to accurately assess the level of transmitted drug resistance in Cuba, as the extent of drug resistance might jeopardize effectiveness of first-line regimens prescribed in Cuba and might necessitate the implementation of baseline drug resistance testing.

High frequency of antiviral drug resistance and non-B subtypes in HIV-1 patients failing antiviral therapy in Cuba.

BACKGROUND: Emergence of HIV-1 drug resistance may limit the sustained benefits of antiretroviral therapy (ART) in settings with limited laboratory monitoring and drug options. OBJECTIVES: Surveillance of drug resistance and subtypes in HIV-1 patients failing ART in Cuba. STUDY DESIGN: This study compiled data of ART-experienced HIV-1 patients attending a clinical center in Havana in 2003 and 2009-2011. The first period included results of a cross-sectional study, whereas in the second period genotyping was performed as part of routine care. Drug resistance mutations and levels were determined using HIVdb version 6.0.9. RESULTS: Seventy-six percent received solely ART containing at least 3 drugs, of which 79.1% ever receiving unboosted protease inhibitors (PI). Patients from 2009 to 2011 were longer treated and exposed to more ART regimens. Subtype B (39%) and CRF19_cpx (18%) were the most prevalent genetic forms. Subtype distribution did not change significantly between both periods, except for BG recombinants that increased from 6% to 14%. Nucleoside reverse transcriptase inhibitor (NRTI), non-nucleoside RTI (NNRTI) and PI mutations were present in 69.5%, 54.8% and 44.4%. Full-class resistance (FCR) to NRTI, NNRTI, PI and multidrug resistance (MDR) were detected in 31.8%, 37.9%, 18.5% and 15.4%. FCR to NRTI, NNRTI, PI and MDR were present in 9.8%, 14.1%, 0%, 0% after first-line failure and in 19.8%, 20.8%, 2.9% and 2.9% after second-line failure. CONCLUSIONS: Our study found a high prevalence of drug resistance and supports the need for appropriate laboratory monitoring in clinical practice and access to drug options in case of virological failure.