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Introduction: Genital Chlamydia trachomatis infection is one of the most frequent in the world; about 85 million new cases of this pathology are registered each year, which causes severe complications in women and newborns. Objective: To determine the clinical-epidemiological characteristics of C. trachomatis infection in sexually active Venezuelan women. Materials and methods: Descriptive, cross-sectional, and field study based on the clinical history and physical examination, detection of infection with immunoenzymatic assay with anti- LPS polyclonal antibody and confirmation of results with molecular biology test. The sample consisted of 100 sexually active women over 12 years of age from Carabobo state, Venezuela. Results: The women were mostly between 20 and 45 years old, in 25% of them IgG antibodies to C. trachomatis were detected and in 84% of these the infection was confirmed by PCR, in none of the women IgM antibodies to C. trachomatis were found. Conclusion: Chronic infection characterizes women between 20 and 45 years of age; the immunoenzymatic test yielded false positives corroborated by PCR. [ABSTRACT FROM AUTHOR]

Introduction: Toxoplasma gondii is a parasite with great zoonotic potential. It can infect a broad range of warm-blooded hosts (including livestock) and causes significant losses in the industry. In humans, it has been described as a pathogen in immunosuppressed people, it affects the fetus development in congenital infections, and is associated with various behavioral disorders in healthy people. Humans can acquire T. gondii by consuming undercooked, contaminated meat. Objective: To determine T. gondii positivity (currently unknown) in meat for human consumption (i.e., beef, chicken, and pork) in the city of Ibague, Colombia. Materials and methods: We used conventional nested PCR and the T. gondii B1 gene sequence as amplification target. We collected samples of meat (N=186) sold in the urban area of Ibagué (62 beef, 62 chicken, and 62 pork samples) and determined the T. gondii positivity percentage for each type of meat. Results: The study found an average of 18.8% positivity for all meat samples, pork having the highest percentage (22.5%; 14/62), followed by beef (19.3%; 12/62) and chicken (14.5%; 9/62). The best-amplified products were sequenced by macrogen and aligned with the B1 gene sequences in GenBank, thereby confirming their identity. Conclusions: This is the first study of T. gondii prevalence in meat for human consumption carried out in the city of Ibagué and the department of Tolima. All three types of meat sampled represent a risk for local human infection. [ABSTRACT FROM AUTHOR]

Introduction: Bats have been reported as hosts of the Trypanosoma cruzi protozoan, the etiologic agent of American trypanosomiasis, an endemic zoonotic disease in México. Objective: To describe T. cruzi infection in bats from the states of Campeche and Yucatán, México. Materials and methods: Captures were made from March to November, 2017, at three sites in Yucatán and one in Campeche. Up to four mist nets on two consecutive nights were used for the capture. The bats' species were identified and euthanasia was performed to collect kidney and heart samples for total DNA extraction. Trypanosoma cruzi infection was detected by conventional PCR with the amplification of a fragment belonging to the T. cruzi DNA nuclear. Results: Eighty-six bats belonging to five families (Vespertilionidae, Noctilionidae, Mormoopidae, Phyllostomidae, and Molossidae) and 13 species (Rhogeessa aeneus, Noctilio leporinus, Pteronotus davyi, P. parnellii, Artibeus jamaicensis, A. lituratus, A. phaeotis, Glossophaga soricina, Carollia sowelli, Chiroderma villosum, Uroderma bilobatum, Sturnira parvidens, and Molossus rufus) were captured. Infection frequency by PCR was 30,2% (26/86) detected only in the renal tissue. The infected species were P. parnellii, G. soricina, A. lituratus, A. jamaicensis, S. parvidens, C. villosum, and R. aeneus. Conclusions: Our results confirmed the participation of several bat species as hosts in the T. cruzi transmission cycle in the region. Further studies are necessary to establish the importance of these animals in the zoonotic transmission of T. cruzi. [ABSTRACT FROM AUTHOR]

Introduction: Toxoplasmosis is an orally-transmitted zoonosis that may appear after consuming food contaminated with any infective form of Toxoplasma gondii. Its transmission by water has been reported in several countries including Colombia. The rural population of Sincelejo could be at risk of contracting toxoplasmosis through this route given that they lack potable water. Objective: To evaluate T. gondii contamination in water for human consumption from water wells (jagüeyes) in the rural area of Sincelejo and establish its relationship with different social determinants of health in the study area. Materials and methods: Using nested PCR we evaluated 96 water samples obtained from 48 farms located in eight rural townships in Sincelejo. We took two samples in each farm: one of raw water from water wells and the other intended for direct consumption. We conducted a survey on each farm to collect information on the physical characteristics of dwellings, the presence of cats, and the availability and uses of water. Statistical relationships were evaluated through Fisher tests. Results: Of the 96 samples analyzed, 13 were contaminated with T. gondii (13.5%): Nine corresponded to raw water and four to water for direct consumption. No statistical association was found between the positive samples and the social determinants of health under evaluation (p>0.05). Conclusion: The rural population of Sincelejo could be at risk of contracting toxoplasmosis through the use and/or consumption of water from its water wells. The contamination of these water bodies by T. gondii may be influenced by unstudied social determinants. [ABSTRACT FROM AUTHOR]

Introduction: Chagas' disease is the leading cause of infectious myocarditis worldwide. This infection caused by Trypanosoma cruzi is usually life-long and asymptomatic; however, the third part of infected people can develop severe or even fatal cardiomyopathy. As the parasitemia in the chronic phase is both low-grade and intermittent, T. cruzi infection is principally detected by serology, although this method has sensitivity and specificity limitations. Objective: To determine the level of agreement between serologic and molecular tests in 658 voluntary blood donors from six provinces in the Colombian department of Santander. Materials and methods: We evaluated an array of diagnostic technologies by cross-section sampling performing a serological double diagnostic test for T. cruzi antibody detection (Chagas III ELISA™, BiosChile Group, and ARCHITECT Chagas CMIA™, Abbott), and DNA detection by polymerase chain reaction (PCR). We collected the demographic, clinical, and epidemiological information of participants. The sample size was calculated using Epidat™ and the statistical analysis was done with Stata 12.1™. Results: PCR was six times more sensitive in detecting T. cruzi infection than ELISA/CMIA with prevalence values of 1.8% (12/658) and 0.3% (2/658), respectively, and kappa=0.28 (95%CI: -0.03 - 0.59). In contrast, serology showed a sensitivity of 16.7% (95%CI: 2.09 - 48.4) and a specificity of 100% (95%CI: 99.4 - 100). All seropositive samples were found to be positive by PCR. Conclusions: The implementation of PCR as a complementary method for screening donors could reduce the probability of false negative and the consequent risk of transfusional-transmission of Chagas' disease, especially in endemic regions. [ABSTRACT FROM AUTHOR]

Introduction: In Nicaragua, it is required to standardize molecular tests such as real-time PCR (qPCR) that improve the diagnosis of leptospirosis in humans and animals. Objective: To evaluate two qPCR for detection of pathogenic leptospires in domestic animals of Nicaragua. Materials and methods: Primers were designed for the amplification of the LipL32 gene in SYBR Green (SYBR Green-A in this study) and TaqMan, in addition, previously described primers were used (SYBR Green-B, in this study). The sequences of 12 strains obtained from the database of the National Center for Biotechnology Information (NCBI) were aligned to search for probes and primers. Analytical sensitivity was determined by calculating the detectable genomic equivalent (EG), while 18 pathogenic references strains and 28 negative controls were used in the diagnostic sensitivity and specificity respectively, the methods were applied in 129 urine samples of domestic animals Results: In SYBR green-A, got a detection limit of 4 EG per reaction, in TaqMan, sensitivity was 94.4% (IC95% = 81.1-100.0), in SYBR Green-A obtained sensitivity of 77.8% (IC95% = 55.8-99.8), while SYBR Green-B 61.1% (95%CI = 35.8-86.4). In the three tests, a specificity of 100% (95% CI = 98.2-100.0) was obtained. In field samples 26.4% were positive with SYBR Green-A and 6.1% with SYBR Green-B. Conclusion: The SYBR green-A presented a low detection limit, while the three evaluated techniques showed high specificity, but the TaqMan presented the highest sensitivity. [ABSTRACT FROM AUTHOR]

San Andrés and Providencia are Colombian islands in the Caribbean Sea. San Andrés has 68,283 inhabitants and has registered cases of leprosy in immigrants from continental Colombia. Providencia has 5037 inhabitants and historically health programs do not have records of the disease, but in 2009 two cases of multibacillary histoid leprosy have been confirmed and subsequently another two, representing a prevalence of 8 cases per 10,000 inhabitants, which qualify this site as hyperendemic for leprosy. Initially a 14-year-old girl with histioid leprosy was diagnosed and, exploring this case, her father was diagnosed with the same clinical leprosy form. Recently, a new intrafamilial patient with multibacillary leprosy and an extra-familial girl with undetermined leprosy were detected. The objective of this work is to present to the scientific community and public health entities these clinical cases and to draw the attention of the sanitary authorities on the necessity of establishing continuous programs of epidemiological surveillance of the leprosy in the island using the new tools available in the Leprosy Control Program. [ABSTRACT FROM AUTHOR]

Introduction: Taking into account the difficulty of performing microscopic diagnosis in rural areas, PDRs are a good alternative for the diagnosis of malaria, so it is important to know their diagnostic performance. Objective: To evaluate the diagnostic performance of the PDRs used in 5 departments against the microscopic diagnosis, using PCR as a reference standard. Materials and methods: Thick blood film and PDR were taken to symptomatic individuals who came for the diagnosis of malaria and additionally a filter paper with blood was impregnated for the molecular test. Results: It was included 314 samples whose percentage of positivity for malaria was 49 % by PCR, 48% by microscopy and 46% by PDR, with parasitemias between 180 p/μL and 23,800 p/μL of blood. The concordance of microscopy results (PTO MIC) versus PCR (LNR) was: Cohen kappa coefficient 0.975 (95% CI: 0.950-0.999), sensitivity 97% (95% CI 95-100) and 100% specificity (95% CI: 100-100), kappa index of species 0.958 (IC95%: 0.912-1.00). The concordance between PDR Pf/Pv (PTO MIC) versus PCR (LNR) was: kappa coefficient of 0.878 (95% CI: 0.784-0.973), sensitivity 94% (95% CI: 87-100), 95% specificity (95% CI: 90-100), kappa index of species 1.0 (95% CI: 1.00-1.00). The concordance between PDR Pf/Pan versus PCR was: Cohen's kappa coefficient of 0.920 (95 % CI: 0.865- 0.974), sensitivity 94% (95% CI: 90-98), 99% specificity (95% CI 95- 100), kappa index of species 0,750 (IC95% 0,637-0,863). Conclusion: The results of this study support the use of PDR in Colombia; however, it requires more training of personnel in the use of PDR and use PDR where the Plasmodium species can easily be differentiated. [ABSTRACT FROM AUTHOR]

Introduction: Mucosal leishmaniasis has a progressive course and can cause deformity and even mutilation in the affected areas. It is endemic in the American continent and is mainly caused by Leishmania (Viannia) braziliensis. Objective: To describe a series of mucosal leishmaniasis cases and the infectious Leishmania species. Materials and methods: Fifty patients with a clinical diagnosis of mucosal leishmaniasis and parasitological confirmation were included. We describe the clinical and laboratory results. Species typing was performed by PCR-RFLP using miniexon and hsp70 genes and confirmed by sequencing. Results: The median time of disease evolution was 2.9 years (1 month to 16 years). The relevant clinical findings included mucosal infiltration (94%), cutaneous leishmaniasis scar (74%), total loss of the nasal septum (24%), nasal deformity (22%) and mucosal ulceration (38%). The symptoms reported included nasal obstruction (90%), epistaxis (72%), rhinorrhea (72%), dysphonia (28%), dysphagia (18%) and nasal pruritus (34%). Histopathology revealed a pattern compatible with leishmaniasis in 86% of the biopsies, and amastigotes were identified in 14%. The Montenegro skin test was positive in 86% of patients, immunofluorescence in 84%, and culture in 8%. L. (V.) braziliensis was identified in 88% of the samples, L. (V) panamensis in 8%, L. (V.) guyanensis and L. (L.) amazonensis in 2%. Conclusion: In this study, we found a severe nasal disease with destruction and deformity of the nasal septum, in a quarter of the cases that was probably related to late diagnosis. L. (V.) braziliensis was the predominant species. A case of mucosal leishmaniasis in Colombia caused by L. (L.) amazonensis is described for the first time. [ABSTRACT FROM AUTHOR]

Introduction: Cutaneous leishmaniasis caused by L. braziliensis has been historically endemic in Argentina is an historical endemic area and several cases of visceral leishmaniasis following previous cutaneous leishmaniasis have been reported. Currently, visceral leishmaniasis affecting humans and dogs is being found in the city of Posadas (Province of Misiones, Argentina) since 2006. Objective: Identify the etiologic agent to species level in patients with visceral leishmaniasis diagnosis in Misiones province and describe its clinical and epidemiological characteristics. Materials and methods: Twenty-four patients from Misiones province were studied from 2009 to 2016, all with a confirmed diagnosis of visceral leishmaniasis. To identify the Leishmania species involved, patient samples were analyzed by microscopy, serologic studies, DNA detection and sequencing. Variables like age, sex, clinical signs and symptoms consistent with visceral leishmaniasis were recorded. Results: 75% (18/24) of the patients studied were males and 25% (6/24) were younger than 4 years. The most frequent symptom prolonged fever in 87.5% of the patients (21/24), followed by splenomegaly in 70.8% (17/24). Leishmania infantum was the only parasite species identified in all patients. Conclusion: This finding constitutes the first molecular identification of Leishmania infantum species in autochthonous patients of Misiones province, Argentina. This study highlights the importance of the PCR for species identification in epidemiological studies of visceral leishmaniosis in Argentina. [ABSTRACT FROM AUTHOR]

Introduction: The study of the interaction between the parasite, the vector and the mammalian hosts, including man, allows to understand the behavior of the leishmaniases. Objetive: To determine the presence of Lutzomyia species and to detect the Leishmania infection in D. marsupialis in an endemic area for visceral leishmaniasis. Materials and methods: Phlebotomine fauna and individuals of D. marsupialis were collected with CDC and Tomahawk® traps, respectively. The species of Lutzomyia were identified using the Young & Duncan (1994) key. Ear and tail biopsies and blood samples from D. marsupialis were taken to identify the Leishmania species by amplifying a fragment of the gene associated with the 70 kD heat shock protein. Results: Seven Lutzomyia species were identified: Lu. evansi, Lu. gomezi, Lu. panamensis, Lu. dubitans, Lu. cayennensis cayennensis, Lu. rangeliana and Lu. trinidadensis. The first three species have epidemiological importance in Colombia because of their implications in the transmission of the Leishmania parasite. Sixty-five tissue samples from 19 D. marsupialis individuals were negative for Leishmania spp. Conclusions: The presence of the Lutzomyia species that have been identified as vectors for Leishmania inside and around houses in the village of El Bledo, in El Carmen de Bolivar represents a risk of infection. Furthermore, the presence of Lu. panamensis is reported for first time in El Carmen de Bolivar in Colombia. Although the lack of detection of Leishmania spp in D. marsupialis samples may suggest that D. marsupialis does not play an important role in the transmission cycle of Leishmania in this region, it is necessary to carry out further longitudinal studies to confirm this hypothesis. [ABSTRACT FROM AUTHOR]

Introduction: Police working horses live in close contact with their handlers and the general population during recreational and patrol activities, an aspect that may favor the transmission of leptospirosis between horses and occupationally personnel. Objective: To characterize the epidemiology of leptospirosis by serology, uroculture and PCR in working horses and population with occupational risk of six Colombian police units. Material and methods: A total of 153 castrated male horses and 123 people from six police units of Manizales, Pereira, Armenia, Ibagué, Tuluá and Cali municipalities were evaluated. Three structured instruments were applied and blood samples from people and horses were obtained, which were processed by MAT with 24 serogroups. A clinical examen of horses was done and urine samples were obtained for urine culture and a conventional PCR. Results: The seroprevalence of human leptospira spp. was 3.25 % (n=4) and 85 % (n=130) in horses. The serogroups L. Djasiman y L. Shermani were the most prevalent in horses. The urine culture was positive in 64.7 % (99/153) of the samples, whereas the PCR analyzes were negative. A statistically significant association was found in the frequency of departure from the facilities (P=0.009) and the presence of wildlife (P=0.051), with infection by L. Shermani serogroup. Conclusion: The epidemiological characteristics of horse leptospirosis suggest an endemic presentation of the infection, and the role of horses as reservoirs of the bacteria, but it is necessary to elucidate the pathogenesis of the disease with complementary studies. [ABSTRACT FROM AUTHOR]

Introduction: Each year approximately 3 million people die as the result of foodborne diseases. The fresh artisan (handmade) cheese produced and distributed in the Colombian Caribbean region is a native product from the departments of Córdoba, Sucre, Bolívar, Atlántico, Magdalena, Cesar, and La Guajira. Its mass consumption increases the risk of infection with Salmonella spp., Listeria spp., and Brucella spp., as it is made with a very rustic technology, with unpasteurized cow milk, without standardized and hygienic procedures and its storage is inadequate. Objective: To detect the presence of Salmonella spp., Listeria spp., and Brucella spp. in samples of fresh artisan cheese from the Colombian Caribbean region. Materials and methods: Twenty-seven samples of cheese from five departments of the Caribbean Region (Atlántico (n=6), Bolívar (n=2), Córdoba (n=1), Magdalena (n=16), and Sucre (n=2)) were analyzed by real-time polymerase chain reaction (qPCR). Seventeen of the samples corresponded to soft cheese, five to semi-hard cheese and five to hard cheese. Results: In 62.9% (17/27) of the samples we detected Salmonella spp., in 70.4% (19/27), Listeria spp., and in 22.2% (6/27), Brucella spp., mainly from the department of Magdalena. In 62.5% (10/16) of the samples we detected Salmonella spp. and Listeria spp. while in the department of Atlántico, 50% (3/6) of the samples corresponded to Brucella spp. Conclusion: The results confirmed the presence of these microorganisms in all the samples of soft cheese from the Colombian Caribbean region. [ABSTRACT FROM AUTHOR]

Introduction: Candidiases is a group of opportunistic infections caused by yeasts belonging to the genus Candida. Candida albicans is the most prevalent species in both superficial and deep infections, however, the clinical importance of non-albicans Candida has increased during the last decade, driving an urgent need for diagnostic tests that allow for species-level resolution and selection of the optimum therapeutic approach. Objective: To design and to optimize a new multiplex PCR assay for the simultaneous identification of the five most relevant species of Candida involved in human candidiasis etiology. Materials and methods: For primers design, the physical and thermodynamic restrictions that affect multiplex PCR performance were analyzed using Gene Runner and Mult-PSOS. As templates, the internal transcribed region 2 (ITR2) was selected for C. albicans (AJ249486.1), and topoisomerase II (TOPII) for C. parasilopsis (AB049144.1), C. krusei (AB049139.1), C. tropicalis (AB049141.1), and C. guillermondii (AB049145.1). We used ATCC strains of all these five species and clinical isolates as templates. Results: We designed ten oligonucleotides for the simultaneous amplification of the Candida species. The electrophoresis band profile was: C. albicans (206 bp), C. guillermondii (244 bp), C. tropicalis (474 bp), C. parasilopsis (558 bp), and C. krusei (419 bp). Conclusion: The new multiplex PCR assay designed in this study allowed a simultaneous and efficient amplification of the amplicons corresponding to the five species of Candida under study, with an adequate resolution in standard agarose gel. [ABSTRACT FROM AUTHOR]

Introduction: There are few reports from Venezuela describing the genetic basis that sustains the pathogenic potential and phylogenetics of Escherichia coli extraintestinal strains isolated in health care units. Objective: To establish the genetic diversity of extraintestinal E. coli strains producers of betalactamases TEM, SHV and CTX-M associated with healthcare. Materials and methods: We studied a collection of 12 strains of extraintestinal E. coli with diminished sensitivity to broad-spectrum cephalosporins. Antimicrobial susceptibility was determined by minimum inhibitory concentration. We determined the phylogenetic groups, virulence factors and genes encoding antimicrobial resistance using PCR, and clonal characterization by repetitive element palindromic-PCR rep-PCR. Results: All strains showed resistance to cephalosporins and joint resistance to quinolones and aminoglycosides. The phylogenetic distribution showed that the A and B1 groups were the most frequent, followed by D and B2. We found all the virulence factors analyzed in the B2 group, and fimH gene was the most frequent among them. We found blaCTX-M in all strains, with a higher prevalence of blaCTX-M-8 two of these strains showed coproduction of bla CTX-M-9 and were genetically identified as bla CTX-M-65 and bla CTX-M-147 by sequencing. Conclusion: The strains under study showed genetic diversity, hosting a variety of virulence genes, as well as antimicrobial resistance with no particular phylogroup prevalence. This is the first report of bla CTX-M alleles in Venezuela and in the world associated to non-genetically related strains isolated in health care units, a situation that deserves attention, as well as the rationalization of antimicrobials use. [ABSTRACT FROM AUTHOR]

Introduction. Difficulties in anopheline identification due to intra-individual variation and interspecies similarity have led to the development of molecular tools that allow species discrimination. Objective. To use bioinformatics tools to develop a PCR-RFLP assay for molecular identification of species belonging to the Oswaldoi Group reported in Colombia and to evaluate the usefulness of a PCR-RFLP-AluI assay previously developed in our lab, in anophelines from different Colombian localities. Material and Methods. In silico restriction patterns were generated by bioinformatics analysis of ITS2 sequences reported in GenBank, corresponding to species belonging to the Oswaldoi Group. To evaluate the PCR-RFLP-AluI assay we used mosquitoes collected in four localities of the Pacific Coast of Colombia: Nuquí, Pizarro, Buenaventura and Tumaco. Results. Bioinformatic analysis of reported ITS2 sequences showed that the enzyme AluI produced in silico restriction patterns that distinguished four species belonging to the Oswaldoi Group; two additional species could be differentiated with a second digest. The evaluation of the PCR- RFLP-AluI confirmed results of morphological identifications and demonstrated identification problems that can arise due to intraspecific variation and interspecies similarity. Conclusions. The expansion of the PCR-RFLP-AluI methodology and its application in different regions of the country will allow rapid and accurate discrimination of a large number of mosquitoes, some of them belonging to species that are difficult to discriminate morphologically. This strategy will be useful for complementary studies to assess malaria transmission dynamics in Colombia. [ABSTRACT FROM AUTHOR]

Introducción. La tuberculosis es la primera causa de muerte por un único agente infeccioso y, la novena, a nivel general en el mundo. Su agente etiológico son especies del complejo Mycobacterium tuberculosis, al cual pertenece Mycobacterium bovis, agente de transmisión zoonótica. Para el año 2017, la Organización Mundial de la Salud (OMS) estimó 142.000 casos nuevos en humanos de tuberculosis zoonótica y 12.500 muertes. En Colombia en la última década, se reportó un caso de tuberculosis humana por esta especie, situación por investigar teniendo en cuenta que el Instituto Colombiano Agropecuario (ICA) reporta el sacrificio regular de bovinos con tuberculosis confirmada de diferentes departamentos. Se buscó identificar casos de tuberculosis producidos por M. bovis a partir del estudio de aislamientos clínicos que se recuperan por la vigilancia realizada en el Laboratorio Nacional de Referencia del Instituto Nacional de Salud. Materiales y métodos. Se seleccionaron 221 aislamientos clínicos por sus características de resistencia y presencia de infección concomitante con HIV. Se usó la PCR multiplex en tiempo real, para identificar las especies M. tuberculosis, M. bovis y M. bovis BCG, y se describió el perfil de sensibilidad a los fármacos. Resultados. El 18 % (n=39) de los aislamientos fueron resistentes a pirazinamida, el 5,4 % (n=12) resultaron multirresistentes y el 7,7 % (n=17) fueron monorresistentes a rifampicina; 72 aislamientos pertenecían a pacientes con coinfección por HIV. Todos los aislamientos se identificaron como M. tuberculosis, a excepción de uno que fue clasificado como no perteneciente al complejo. Ningún aislamiento resultó ser M. bovis. Conclusión. Mediante los aislamientos estudiados, no fue posible demostrar la presencia de tuberculosis por M. bovis, resultados concordantes con lo reportado previamente en el país (4,5). [ABSTRACT FROM AUTHOR]

Introduction: Viral encephalitis is a well-known inflammatory process associated with neurological dysfunction that might derive into severe brain damage or a fatal outcome. In México there is no epidemiological data that describes the prevalence of viral agents responsible for acute encephalitis. Objective: To identify the main viral agents by real time PCR involved in acute encephalitis in Mexico. Materials and methods: We obtained cerebral spinal fluid (CSF) samples from all patients with suspected viral encephalitis admitted to the emergency service of the Hospital Civil de Guadalajara "Fray Antonio Alcalde". To identify pathogens, we performed nucleic acid extraction using real-time PCR and RT-PCR. Results: Sixty-six patients were diagnosed with acute encephalitis from 2011 to 2014. A definitive viral etiological diagnosis was established in 16 patients (24%); the main causative agents were enteroviruses in 50% of the 16 positive samples, followed by herpes simplex virus (37%) and cytomegaloviruses (12.5%). Patients with encephalitis were predominantly male (63.3%) and a seasonal predominance was observed during autumn (37.5%). The main clinical characteristics in the acute encephalitis phase were fever (48.45) and cephalea (36.3), followed by seizures, disorientation, and muscular weakness (30.3%). Kerning sign was present in two cases (3%) and other two cases presented Brudzinski's sign (3%). Conclusions: CSF PCR is a suitable diagnostic technique for the identification of viral encephalitis caused by viral infections that allows an appropriate antiviral therapeutic treatment. [ABSTRACT FROM AUTHOR]

Introduction: Syphilis is a disease produced by Treponema pallidum subspecies pallidum, which affects approximately 12 million people worldwide every year. Of these, more than 2 million are pregnant women whose babies end up having congenital syphilis, the worst form of this infection. Objective: To detect the presence of T. pallidum subspecies pallidum in clinical samples in order to diagnose congenital syphilis by means of nested PCR, and to determine its concordance with serological testing. Materials and methods: Three target genes (polA, 16S ADNr y TpN47) were amplified by conventional and nested PCR. The results from the amplification of the TpN47 and polA genes were confirmed by sequencing. The serological tests used were VDRL (Venereal Disease Research Laboratory), RPR (Rapid Plasma Reagin) y TPPA (Treponema pallidum Particle Agglutination Assay). Results: The sensitivity for the conventional PCR was 52 pg and 0.52 pg for the nested PCR. The specificity of primers TpN47 and polA was 100%; the results of the sequencing showed a 97% identity with T. pallidum. There was concordance between the serology and the nested PCR in 70% of the samples. Conclusion: The TpN47 gene was the best molecular target for the identification of T. pallidum. The nested PCR is a promising molecular tool for the diagnosis of congenital syphilis. [ABSTRACT FROM AUTHOR]

Introduction: Multilocus enzyme electrophoresis (MLEE) is the reference standard for the characterization of Leishmania species. The test is restricted to specialized laboratories due to its technical complexity, cost, and time required to obtain results. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is used to identify Leishmania species. Objective: To establish the concordance between the two tests as identifying methods for circulating species in Colombia. Materials and methods: A total of 96 isolates from patients with cutaneous or mucosal leishmaniasis were selected and identified by MLEE and PCR-RFLP with miniexon and hsp70 as the molecular targets, which were used sequentially. Restriction enzymes HaeIII and BccI were similarly applied. Cohen's kappa coefficient and the 95% confidence interval (CI) were calculated. Results: The kappa coefficient and the 95% CI between MLEE and PCR-RFLP displayed "very good" concordance with a coefficient of 0.98 (CI95%: 0.98 to 1.00). The identified species were Leishmania Viannia braziliensis, Leishmania Viannia panamensis, Leishmania Viannia guyanensis and Leishmania Leishmania amazonensis. A total of 80 of the 96 isolates were sequenced and the results obtained by PCR-RFLP were confirmed. Conclusion: Due to the concordance obtained between tests results with the amplification of the genes miniexon and hsp70, PCR-RFLP is proposed as an alternative for identifying circulating Leishmania species in Colombia. [ABSTRACT FROM AUTHOR]

Introduction: The genetic makeup of the host contributes to the clinical profile of dengue. This could be due to the effect of variants in the genes encoding pro-inflammatory cytokines. Objective: To evaluate the association between the variants of three polymorphisms in TNFA, IL6 and IFNG candidate genes with dengue severity in a sample of Colombian population. Materials and methods: We evaluated the rs1800750, rs2069843, and rs2069705 polymorphisms in TNFA, IL6 and IFNG candidate genes, respectively, in 226 patients with dengue infection. The genotypes were typed using both polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). To determine the risk of different dengue phenotypes, we compared allele frequencies with chisquare and genotypes and haplotypes using logistic regression. Finally, these analyzes were adjusted with data from self-identification or the ancestral genetic component. Results: The A allele in the rs2069843 polymorphism, adjusted by self-identification, was associated with dengue hemorrhagic fever cases in Afro-Colombians. In the entire sample, this polymorphism, adjusted by the ancestral genetic component, was reproducible. In addition, there were significant associations between GGT and GAC allelic combinations of rs1800750, rs2069843, and rs2069705 in dengue hemorrhagic fever patients, with and without adjustment by ancestral genetic component. Additionally, the AGC allelic combination produced 58.03 pg/ml of interleukin-6 more than the GGC combination, regardless of European, Amerindian and African genetic components. Conclusions: The variants of GGT and GAC polymorphisms of rs1800750, rs2069843, and rs2069705 in the TNFA, IL6 and IFNG genes, respectively, were correlated with the susceptibility to dengue severity in a sample of Colombian population. [ABSTRACT FROM AUTHOR]

Introduction: Clostridium difficile is the main pathogen related to healthcare-associated diarrhea and it is the cause of 20 to 30% of diarrhea cases caused by antibiotics. In Colombia and Latin America, the knowledge about the epidemiological behavior of this infection is limited. Objective: To describe the characteristics of a series of patients with C. difficile infection. Materials and methods: We performed a descriptive case series study of patients with C. difficile infection hospitalized in the Fundación Clínica Shaio from January, 2012, to November, 2015. Results: We analyzed 36 patients. The average age was 65 years. The risk factors associated with the infection were: previous use of antibiotics (94.4%), prior hospitalization in the last three months (66.7%) and use of proton pump inhibitors (50%). The most common comorbidities were chronic kidney disease (41.7%) and diabetes mellitus (30.6%). The most frequent symptoms were more than three loose stools per day (97.1%) and abdominal pain (42.9%). According to the severity of the disease, 44.4% of cases were classified as mild to moderate, 38.9% as severe, and 11.1% as complicated or severe. The detection of the toxin by PCR (GeneXpert) was the most common diagnostic procedure (63.8%). Global mortality during hospitalization was 8%. We identified four strains with serotype NAP1/027 and nine samples positive for binary toxin. Conclusion: Clostridium difficile infection should be suspected in patients with diarrhea and traditional risk factors associated with this disease. We report the circulation of the hypervirulent strain serotype NAP1/027 in Colombia, which should be countered with epidemiological surveillance and a prompt diagnosis. [ABSTRACT FROM AUTHOR]

Introduction: Cutaneous leishmaniasis, caused by parasites of the genus Leishmania, is a disease with high incidence in Colombia. The diagnosis and identification of the infectious species are critical factors when selecting and initiating treatment. Currently, the methods for diagnosing and typing cutaneous leishmaniasis require complicated procedures and there is a need for the validation of new molecular markers and methods to simplify the process. Objective: To develop a tool based in PCR melting curves (PCR-HRM) for the diagnosis and typing of the three Leishmania species of epidemiological importance for cutaneous leishmaniasis in Colombia. Materials and methods: The genomes of Leishmania panamensis, L. braziliensis and L. guyanensis were compared with bioinformatic methods. The species-specific regions were then validated using PCR. For the selected markers, a PCR-HRM was designed, and validity and security parameters were estimated using isolates from Colombian patients previously characterized by PCR-RFLP of the hsp70 gene. Results: The comparative genomic analysis yielded 24 species-specific regions. However, the PCR validation identified only one marker that was specific to each Leishmania species. The other markers showed cross amplification. The detection limit for the three selected markers was one parasite. The sensitivity, specificity, predictive positive and negative values were 91.4%, 100%, 100% and 75%, respectively. Conclusions: The three selected regions can be used as molecular markers in the diagnosis and typing of the causative species of cutaneous leishmaniasis in Colombia. [ABSTRACT FROM AUTHOR]

Introduction: Chronic hepatitis C virus infection is a worldwide public health problem; it has been estimated that over 180 million people are infected with this virus worldwide. Its precise incidence and prevalence (i.e., epidemiology) and the most frequent circulating genotypes in Colombia are unknown. Objective: To describe the hepatitis C virus (HCV) genotypes and subtypes in infected Colombian patients. Materials and methods: We recovered the data on 1,538 HCV isolates from 1,527 patients in two Colombian reference laboratories typed by PAGE or qPCR. Results: Patients' mean age was 53 years; 70% of them were 40 to 70 years old, and 52%, females; 57% of all tests were ordered in Bogotá and 80% of cases were from Cundinamarca, Valle and Atlántico departments. Genotype 1 was detected in 88.6% of cases, distributed as follows: 70% subtype 1b, 13.5% subtype 1a and 5.1%, undetermined subtypes. Genotype 2 was found in 5.4% of the patients, genotype 3 in 2% and genotype 4 in 4%. Mixed genotypes were found in 0.8% of the samples. Conclusion: Genotype 1 is the most common HCV genotype circulating in Colombia, and subtype 1b the most frequent. [ABSTRACT FROM AUTHOR]

Introduction: The province of Pichincha in Ecuador is an endemic area of cutaneous leishmaniasis, where anthropophilic sand flies with natural infection by Leishmania, have been reported as vectors. However, the role in transmission of zoophilic species has not been evaluated.. Objective: To evaluate natural infection by Leishmania in two zoophilic phlebotomine sand fly species, Lutzomyia reburra and Lu. barrettoi majuscula, and one anthropophilic species, Lu. trapidoi, as well as the endophagy and synanthropism of these species in the northwest of Pichincha. Materials and methods: Phlebotomines were collected using CDC light traps in different habitats and altitudes with presence of cutaneous leishmaniasis. Leishmania infection was detected using genomic DNA from females of the collected sand flies. We amplified the internal transcribed spacer gene of ribosomal RNA I (ITS1), the mitochondrial topoisomerase II gene (mtTOPOII), and the nuclear topoisomerase II gene (TopoII). Percentages of positivity for Leishmania, at spatio-temporal scale, proportion of endophagy and synanthropism index were calculated. Results: Natural infection was determined for Le. amazonensis in Lu. reburra (9.5%) and Lu. b.majuscula (23.8%), while in Lu. trapidoi we detected Le. amazonensis, Le. brazilienis and Le. naiffilainsoni. Phlebotomines were asynanthropic and with low endophagy. Conclusion: Natural infection with Le. amazonensis was recorded for the first time in Lu. reburra and Lu. b. majuscula, demonstrating the importance of zoophilic phlebotomines in the maintenance of the Leishmania transmission cycle in endemic foci. [ABSTRACT FROM AUTHOR]