Infección congénita por citomegalovirus en neonatos muy bajo peso / Congenital cytomegalovirus infection in very low birth weight neonates

Introducción: La infección congénita por el citomegalovirus en neonatos menores de 1500 gramos puede ser causa de morbilidad, mortalidad y discapacidad. Objetivo: Describir el comportamiento de la infección congénita por citomegalovirus en un servicio de neonatología. Métodos: Se realizó un estudio descriptivo y transversal con 61 neonatos. Se les realizó detección de citomegalovirus en la primera semana de vida en suero y orina, mediante reacción en cadena de la polimerasa, para determinar infección congénita. Se evaluaron variables perinatales en todos los neonatos, así como elementos clínicos y resultados de exámenes complementarios en los infectados. Resultados: La incidencia de infección congénita fue de un 10 por ciento (6/61). El 5 por ciento de los estudios fueron positivos (6/122). Ninguna muestra de orina resultó positiva (0/61) y en el 10 por ciento de las muestras de suero (6/61) se detectó el genoma del virus. Se encontró asociación entre valoración nutricional al nacer e infección por citomegalovirus (p< 0,05). El 83 por ciento de los neonatos infectados presentaron algún signo clínico y el síndrome de dificultad respiratoria fue el más frecuente (67 por ciento). En todos los neonatos con infección congénita el ultrasonido cerebral fue normal y en el 33 por ciento se detectó retinopatía de la prematuridad en el fondo de ojo. Conclusiones: La incidencia de infección congénita por citomegalovirus es alta en este grupo de riesgo. Los signos clínicos encontrados y los resultados del fondo de ojo en neonatos con infección congénita se relacionaron con la prematuridad y la valoración nutricional de hipotrófico se asoció con esta infección(AU)

Introduction: Congenital cytomegalovirus infection in neonates weighing less than 1500 grams can be a cause of morbidity, mortality, and disability. Objective: To describe the behavior of congenital cytomegalovirus infection in a neonatal service. Methods: A descriptive and cross-sectional study was conducted with 61 neonates. Cytomegalovirus was detected in the first week of life in serum and urine, by polymerase chain reaction, to determine congenital infection. Perinatal variables were evaluated in all neonates, as well as clinical elements and results of complementary examinations in infected infants. Results: The incidence of congenital infection was 10 percent (6/61). 5 percent of the studies were positive (6/122). No urine samples were positive (0/61) and the virus genome was detected in 10 percent of serum samples (6/61). An association was found between nutritional assessment at birth and cytomegalovirus infection (p < 0.05). A total of 83 percent of infected neonates had some clinical sign, with respiratory distress syndrome being the most common (67 percent). In all neonates with congenital infection, brain ultrasound was normal, and retinopathy of prematurity was detected in 33 percent of patients with fundus retinopathy. Conclusions: The incidence of congenital cytomegalovirus infection is high in this risk group. The clinical signs found and the results of the fundus in neonates with congenital infection were related to prematurity and the nutritional assessment of hypotrophic was associated with this infection(AU)

Evaluación de estuches de PCR-tiempo real para detección de virus del papiloma humano de alto riesgo / Evaluation of real-time PCR kits to detect high-risk human papillomavirus

RESUMEN Introducción: El empleo de técnicas moleculares para el diagnóstico de virus del papiloma humano de alto riesgo oncogénico (VPH-AR) es crucial para la detección precoz del cáncer cervicouterino. Objetivo: Evaluar el desempeño analítico de dos estuches de PCR-tiempo real, comercializados por el Centro de Inmunoensayo de Cuba, para detectar VPH-AR. Métodos: Se utilizaron dos paneles de ADN de muestras cervicouterinas: uno con 150 muestras, para validar el estuche SUMASIGNAL HPV 16/18, el proceso de extracción de ADN y su utilidad como prueba cuantitativa, y otro con 163 muestras para evaluar el estuche HPV 13+2. Se determinó la utilidad clínica del estuche HPV 13+2 en 55 muestras cervicovaginales autocolectadas. Se calcularon los indicadores de desempeño analítico de ambos estuches con respecto a pruebas de referencia. Resultados: Los indicadores de desempeño para SUMASIGNAL HPV 16/18 fueron excelentes (> 95 %), concordancia 96 %, índice kappa=0,93 [0,85-1,01]. La extracción de ADN mostró 100 % de especificidad clínica y analítica y 95 % de sensibilidad analítica. Se obtuvo buena correlación con la prueba de referencia cuantitativa (r = + 0,688). El estuche HPV 13+2 tuvo especificidad y sensibilidad clínicas del 100 %, la especificidad analítica fue del 84 % debido a reactividad cruzada con otros VPH-AR. Su aplicación clínica reveló alta frecuencia de infección (41,8 %): 23,6 % con VPH-AR, particularmente en mujeres jóvenes (50 %). La muestra autocolectada resultó útil (100 %). Conclusión: Los ensayos evaluados mostraron altos estándares de calidad, lo que permitiría su uso con una cobertura nacional en una plataforma tecnológica disponible para todo el país.

ABSTRACT Introduction: The use of molecular techniques for the diagnosis of high oncogenic risk human papillomavirus (hrHPV) is crucial for the early detection of cervical cancer. Objective: To evaluate the analytical performance of two real-time PCR kits, commercialized by the Cuban Immunoassay Center, to detect hrHPV. Methods: Two DNA panels from cervical samples were used: one with 150 samples to validate the SUMASIGNAL HPV 16/18 kit, the DNA extraction process and its usefulness as a quantitative test; and another with 163 samples to evaluate the HPV 13+2 kit. The clinical utility of the HPV 13+2 kit was determined in 55 self-collected cervicovaginal samples. The analytical performance indicators of both kits were calculated with respect to reference tests. Results: Performance indicators for SUMASIGNAL HPV 16/18 were excellent (>95%), concordance 96%, kappa index=0.93 [0.85-1.01]. DNA extraction showed 100% clinical and analytical specificity and 95% analytical sensitivity. Good correlation was obtained with the quantitative reference test (r = + 0.688). The HPV 13+2 kit had 100% clinical specificity and sensitivity, analytical specificity was 84% due to cross-reactivity with other hrHPVs. Its clinical application revealed a high frequency of infection (41.8%): 23.6% with hrHPV, particularly in young women (50%). The self-collected sample was viable (100%). Conclusion: The assays evaluated showed high quality standards, which would allow their use with national coverage in a technological platform available for the whole country.

Subtype-Dependent Co-receptor Tropism in Cuban HIV-1-Infected Patients: Implications for Maraviroc Treatment.

INTRODUCTION: Unlike most high-income countries where subtype B viruses predominate, the Cuban HIV-1 epidemic is characterized by a great diversity of subtypes and circulating recombinant forms. Some studies have shown that HIV variants exhibiting a preference for the CXCR4 co-receptor (X4-tropic) could have impacts on disease pathogenesis, with clinical implications for antiviral treatment plans. Determination of HIV co-receptor tropism is crucial for clinicians in deciding whether maraviroc is an appropriate antiviral. OBJECTIVE: Characterize V3 sequence variability and its relation to viral tropism across different subtypes circulating in Cuba and explore how this may affect treatment success with maraviroc. METHODS: We designed a cross-sectional study that included 72 plasma samples obtained at the Pedro Kourí Tropical Medicine Institute in Havana, Cuba. We sequenced the C2V3 env region and assessed subtype based both on env and pol sequences; tropism was predicted by Geno2pheno analysis. Additionally, 35 V3-loop Cuban sequences, obtained from a previous study, were incorporated into the analysis. Statistical associations among virological, clinical and epidemiological variables were assessed by a chi-square test. RESULTS: Tropism prediction for 72 variants revealed that CRF19_cpx was associated with dual-tropic R5X4 viruses (p = 0.034). Moreover, when 35 sequences from a former study were added, the association was significant not only for R5X4 (p = 0.019) but also for X4-tropic variants (p = 0.044). Alignment of 107 V3-loop sequences showed wide diversity among the different HIV-1 subtypes circulating in Cuba. CONCLUSIONS: In accordance with G2P, CRF19_cpx is a genetic variant with a high proportion of X4 and R5X4-tropic viruses. The results from the present study suggest that the Cuban recombinant could be a more pathogenic variant and that maraviroc may not be suitable for patients infected with CRF19_cpx.

Circulation of Human Papillomavirus and Chlamydia trachomatisin Cuban Women.

INTRODUCTION Human papillomaviruses and Chlamydia tracho-matis are the most frequent causes of sexually transmitted infec-tions. Although the association between some human papillomavirus genotypes and cervical cancer has been demonstrated and Chla-mydia trachomatis infection is the most common cause of female infertility, Cuba has no national baseline studies on the circulation and co-circulation of these agents, the synergistic effect of which may be a risk factor for occurrence and development of precancer-ous cervical lesions. Additionally, few local studies have examined risk factors for infection.OBJECTIVE Determine the frequency of infection by human papil-lomavirus and Chlamydia trachomatis and their association with sociodemographic, clinical and epidemiological variables in women seeking routine Pap smears or other medical services at the primary care level in Cuba.METHODS A cross-sectional study was conducted among 500 wom-en aged 16-67 years (100 from Havana, 200 from Villa Clara and 200 from Holguín Provinces, Cuba), from August through December 2015. Chlamydia trachomatis infection was detected by real-time polymerase chain reaction and 35 genotypes of human papillomavirus by low-density microarray. We then examined the association of infec-tion with sociodemographic, clinical and epidemiological variables.RESULTS Human papillomavirus was detected in 14.8% (74/500) of the women. Of the 29 genotypes identifi ed, 79.7% (59/74) were onco-genic high-risk types. Type 16 was the most frequently identifi ed (23%; 17/74), followed by type 31 (10.8%; 8/74) and then by types 33, 53, 61 and 66 in equal proportions (8.1%; 6/74). Infection frequency was greater in women aged ≤25 years (38.8%; 31/80), students (46.7% 7/15), single women (23.0%; 40/174) and among those who reported having more than 3 sexual partners in the last 2 years (41.5%; 17/41). Differences were found among provinces for circulating genotypes and infection-related variables. Human papillomavirus infection from genotypes 16, 31, 33, 53, 61, 66, 68 and 89 was associated with the 7.9% (30/382) of women who had positive Pap tests. Infection fromChlamydia trachomatis was positive in 1% (5/500) of women, all aged ≤25 years. Coinfection by Chlamydia trachomatis and HPV was found in one woman infected with human papillomavirus genotype 61.CONCLUSIONS Frequency of human papillomavirus is high in the three Cuban provinces studied, with greater frequency of genotype 16 and other oncogenic high-risk types. For both agents, infection is more frequent in young women and adolescents. Positive Pap tests are fre-quently associated with HPV infection. Prevalence fi ndings from this study could be used as a baseline for future research or interventions. KEYWORDS Human papillomavirus, genotypes, Chlamydia tracho-matis, neoplasms, sexually transmitted diseases, cervix Uteri, infec-tion, real-time polymerase chain reaction, women, Cuba.

HIV-1 Antiretroviral Resistance in Cuba, 2009-2014.

INTRODUCTION By the end of 2017, there were more than 28,000 individuals living with HIV in Cuba, over 80% receiving antiretroviral therapy, which dramatically reduces viral replication, improves immune status and decreases risk of transmission. These results could be jeopardized by emergence of HIV-1 drug resistance. In 2009, a test for HIV-1 genotypic resistance was introduced in routine clinical practice in Cuba. OBJECTIVE Investigate antiretroviral resistance and its relation to subtype distribution in HIV-1 treatment-naïve and previously treated patients in Cuba. METHODS Resistance and HIV-1 subtype distribution were determined in 342 antiretroviral treatment-naïve patients and 584 previously treated for HIV-1 whose blood specimens were sent to the Pedro Kourí Tropical Medicine Institute during 2009-2014. Transmitted drug resistance was determined using the Calibrated Population Resistance Tool v.6. Drug resistance analysis was conducted using the algorithm Rega v9.1.0. RESULTS Prevalence of transmitted drug resistance was 11.4%, and 41% of mutated viruses exhibited dual-class resistance to nucleoside reverse transcriptase inhibitor and non-nucleoside reverse transcriptase inhibitor. Overall, 84.9% of patients had ≥1 resistance mutation, 80% had ≥1 nucleoside reverse transcriptase inhibitor mutation, 71.4% had ≥1 non-nucleoside reverse transcriptase inhibitor mutation and 31.7% had ≥1 protease inhibitor mutation. K65R and K101E mutations were significantly more frequent in subtype C, L210W in CRF19_cpx, and M47V/I in CRF BGs (20, 23, 24). Full class resistance to nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, protease inhibitors and multidrug resistance were detected in 21.2%, 32.4%, 8% and 4.1% of patients, respectively. Average percentage resistance to nucleoside reverse transcriptase inhibitor, protease inhibitor, full class resistance to nucleoside reverse transcriptase inhibitor, protease inhibitor and multidrug resistance increased in patients failing two or more regimens. Nevertheless, after 2011, a declining trend was observed in the frequency of multidrug resistance and full class resistance to nucleoside reverse transcriptase inhibitors and protease inhibitors. CONCLUSIONS Detected levels of transmitted drug resistance highlight the need for a national surveillance study in treatment-naïve patients. Resistance prevalence is high in previously treated patients but appears to be decreasing over time. The frequency of resistance mutations in recombinant forms of HIV in Cuba needs further study. KEYWORDS Antiretroviral therapy, highly active antiretroviral therapy, HIV, anti-HIV agents, drug resistance, multiple drug resistance, Cuba.

Detección de anticuerpos IgM contra leptospiras por un sistema comercial ELISA-IgM / Detection of IgM antibodies to leptospires by using a commercial ELISA-IgM system

Introducción: la leptospirosis humana es una zoonosis sistémica y transmisible producida por bacterias invasivas del complejo Leptospira interrogans sensu lato. Numerosos métodos se utilizan para el serodiagnóstico de esta entidad clínica, dentro de los cuales se encuentra ELISA. Objetivos: aplicar un nuevo sistema serológico comercial de ELISA, para la pesquisa de anticuerpos IgM contra leptospiras e identificar los serogrupos de leptospiras presentes en los sueros positivos por este sistema. Métodos: de pacientes sospechosos de leptospirosis se estudiaron 27 y 61 sueros con anticuerpos contra leptospiras y sin estos, respectivamente. La técnica de microaglutinación con antígenos vivos (MAT) se utilizó como método de referencia. Los sistemas comerciales de SD Leptospira ELISA-IgM, SD flujo lateral Leptospira IgM y la hemoaglutinación indirecta (HAT) fueron comparativamente empleados. Resultados: 27 sueros con anticuerpos contra leptospiras resultaron positivos por SD Leptospira ELISA-IgM, 20 por SD flujo lateral Leptospira IgM y 9 por hemoaglutinación indirecta. De 61 sueros sin anticuerpos contra leptospiras, 9 y 8 resultaron positivos, respectivamente, por SD Leptospira ELISA-IgM y por SD flujo lateral Leptospira IgM. Los serogrupos Ballum y Canicola predominaron en los sueros positivos por el sistema comercial. La coincidencia entre SD Leptospira ELISA-IgM y la técnica de microaglutinación con antígenos vivos fue de 89,77 porciento. Conclusiones: SD Leptospira ELISA-IgM muestra una mayor positividad en los sueros estudiados, lo que avalaría su posible introducción en Cuba. Se confirma la amplia reactividad del antígeno usado en SD Leptospira ELISA-IgM, lo cual sugiere mantener una activa vigilancia de laboratorio sobre los serogrupos de leptospiras, a nivel nacional

Introduction: human leptospirosis is a communicable systemic zoonosis caused by the invasive bacteria Leptospira interrogans sensu lato complex. Numerous methods are used for serodiagnosis of this disease, including the ELISA tests. Objectives: to implement a new commercial serological ELISA test (SD Leptospira ELISA-IgM, SD Bioline, Korea) for screening of IgM antibodies and for identification of Leptospira serogroups in positive sera. Methods: twenty seven and sixty one sera, with/without IgM antibodies to leptospires, respectively, were studied. They had been taken from patients suspected of having leptospirosis. The microscopic agglutination test with live antigens (MAT) was the reference method. In addition, other commercial systems such as SD Leptospira IgM Lateral Flow and indirect hemagglutination (HAT) tests were comparatively used. Results: all the 27 sera with antibodies against Leptospira were positive according to SD Leptospira ELISA-IgM, 20 sera were found positive by SD Lateral Flow Igm test and 9 sera by the indirect HAT test. Of the 61 sera without antibodies to leptospires, 9 and 8 were positive by SD Leptospira ELISA-IgM and SD Leptospira IgM Lateral Flow test, respectively. Serogroups Canicola and Ballum were predominant in positive sera tested by the commercial system under evaluation. The agreement coefficient between SD Leptospira ELISA-IgM and MAT was 89.77 percent. Conclusions: SD Leptospira ELISA-IgM showed higher positivity rate than the other systems in the studied sera; this aspect would support its possible introduction in Cuba. The great reactivity of the antigen used in the system was confirmed, which indicates that active laboratory surveillance of leptospiral serogroups should be kept nationwide