A retrospective study of Taenia spp. in Cuban patients: what does molecular analysis tell us?

Taeniosis is a neglected disease, particularly in developing countries, and is caused by infection with the adult tapeworm of either Taenia solium, Taenia saginata, and Taenia asiatica. Of these, T. solium is of primary concern due to the potential for cysticercosis should T. solium eggs be ingested. In Cuba, all cases of taeniosis are assumed to be caused by T. saginata, although some cases of cysticercosis have been documented. It is therefore important to gain further insights regarding the species causing taeniosis in Cuba, especially as diagnostic records indicate an increasing incidence, with the highest number of cases reported in 2020. In this study, we analysed 37 Taenia-positive faecal samples (or proglottids isolated from faecal samples) from the period 2001 until 2020 from all regions of the country. Genomic DNA was extracted from the samples, which had been stored in 10% formalin, using the QIAamp Tissue Kit. Species identification was carried out by duplex real-time PCR targeting the mitochondrial DNA. All cases were found to be T. saginata, and sequence analysis of three isolates confirmed the identification of this species. Our data do not provide any evidence that T. solium currently occurs in Cuba. However, given the relatively low number of samples analysed here, that the parasite may be imported with visitors or travellers who have been in endemic countries, and that taeniosis has relatively mild symptoms and thus infected patients may not seek medical attention, we recommend species determination for all taeniosis cases reported in Cuba.

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.

Herpesviruses excretion in saliva of pediatric transplant recipients.

BACKGROUND: Saliva samples could be used for follow-up of herpesviruses infection in pediatric transplant recipients. OBJECTIVE: With the aim of determining the frequency of herpesviral infections in saliva samples after transplantation, and the association with viremia and complications, a pilot longitudinal follow-up of pediatric Cuban transplanted recipients (kidney and liver) was performed. METHODS: Quantitative real-time polymerase chain reaction of cytomegalovirus (CMV), Epstein-Barr virus, herpes simplex virus, human herpesevirus-6 (HHV6), varicella zoster virus, and human herpesvirus-8 were serially assayed in saliva and serum samples from 27 transplanted patients, during 32 weeks after the graft. Samples taken immediately after the graft were used as control samples. RESULTS: Herpesviruses were detected in 88.9% of saliva and in 37.0% of serum samples. HHV6 and CMV were the viruses more frequently detected (70.4%) in saliva and they were significantly more frequent during the follow-up in comparison with control samples (P < .05). Most patients (22/27) had more than one virus shedding concurrently. Patients with CMV in saliva were associated with CMV viremia (P = .009), particularly at the cutoff of 252.5 copies/mL, with a less accurate level of area under the curve. No association between CMV viral load in saliva and viral disease or response to the antiviral treatment was observed. CONCLUSIONS: The association found between CMV shedding in saliva and CMV viremia in this study opens the possibility of future studies of using viral load in saliva as a predictor of viremia. The implementation of herpesviral load in saliva samples for early clinical intervention in pediatric recipients needs a study with a large number of samples for further conclusions.

Efectos del VIMANG sobre algunos marcadores de progresión de la infección por VIH-1 en pacientes cubanos / Effects of VIMANG on some markers of HIV-1 progression in Cuban patients

Se estudiaron 68 pacientes divididos en 2 grupos, uno que recibió VIMANG durante 6 meses y otro que recibió placebo. Se midieron títulos de anticuerpos anti-p24 y concentración de antígeno p24 al tiempo 0 y 6 meses. Las diferencias encontradas en el comportamiento de los títulos de anticuerpos entre ambos grupos no resultaron ser significativas, aunque se mantuvieron los TPG en el grupo que recibió VIMANG. En el grupo placebo, 2 pacientes elevaron la concentración de antígeno y 2 se mantuvieron positivos para este marcador, mientras que en el grupo estudio desapareció el antígeno en un paciente, que se tornó positivo pero con valores 3 y 4 veces menor que en los 2 pacientes del grupo control, donde este se elevó

Caracterización intratípica de Poliovirus por la técnica de reacción en cadena de la polimerasa

Se introdujo la técnica de la reacción en cadena de la polimerasa para la caracterización intratípica de Poliovirus. Se usaron cebadores que sólo promueven la ampliación de las cepas vacunales de Sabin, comprobada por corrida electroforética de los productos de ADN amplificados (Sabin 1-97 pb, Sabin 2-71 pb, Sabin 3-44 pb) y cuya especificidad se verificó satisfactoriamente. Se estudiaron por esta técnica 23 cepas cubanas de Poliovirus aisladas e identificadas en el Laboratorio de Enterovirus del Instituto de Medicina Tropical "Pedro Kourí" de 1993 a 1994, y todas resultaron ser del tipo vacunal. Se observó cómo el Poliovirus vacunal de Sabin puede ser causa de meningoencefalitis viral como complicación neurológica más leve. Este estudio aportó una evidencia más a favor de la no circulación del Poliovirus salvaje en Cuba.

The polimerase chain reaction techniques was introduced for the intratypic characterization of Poliovirus. Primers were used only to promote the amplification of the Sabin vaccine strains proved by electrophoretic run of the amplified DNA products (Sabin 1 - 97 pb, Sabin 2 - 71 pb, Sabin 3 - 44 pb) and whose specificity was satisfactorily verified. 23 Cuban poliovirus strains isolated and identified at the Laboratory of Enterovirus of the "Pedro Kourí" Tropical Medicine Institute from 1993 to 1994 were studied by this technique. All of them were of the vaccine type. It was observed how the Sabin vaccine poliovirus may be the cause of viral meningoencephalitis as a milder neurological complication. Tghis study provided one more evidence about the non circulation of the wild poliovirus in Cuba.

Estudio de un brote de meningoencefalitis viral: comparación de 2 sistemas biológicos empleados para el aislamiento / Study of an aoutbreak of viral meningoencephalitis: comparison of 2 biological systems used for isolation

Entre los meses de octubre y diciembre de 1995 se reportó un incremento en los casos de meningoencefalitis viral en nuestro país. Se recibieron 23 muestras de heces fecales, que procedían de 2 provincias, con la finalildad de obtener el aislamiento viral mediante la inoculación de éstas en cultivo de células y ratones recién nacidos simultáneamente, para comparar la sensibilidad de ambos sistemas en el aislamiento. Se obtuvo un mayor número de aislamientos en cultivo de células que en ratones, el cultivo de células de riñón de mono verde africano adulto normal (Vero) resultó el más sensible. Se lograron 18 aislamientos, que fueron identificados como Coxsackievirus B5. Se realizó un estudio anatomo-patológico en ratones inoculados y se observaron lesiones compatibles con la infección enteroviral