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Objective: The aim of this study is to differentially identify MAC by PCR in patients with AIDS and disseminated mycobacteriosis., Methods: A cross sectional study was conducted in Mexico to identify MAC by Molecular Biology. Two sets of primers were synthesized: MAV and MIN, for M. avium and M. intracellulare, respectively. Whole-cell DNAs obtained from 29 clinical isolates and clinical serum specimens from other 24 patients with AIDS and disseminated mycobacterial infection were extracted and amplified by PCR with the MAV and MIN primers. The MAV and MIN primers each amplified one highly specific 1.3-kb segment of the homologous DNA, respectively., Results: Twenty-nine DNAs from MAC clinical isolates identified by Gen-Probe AccuProbes were amplified with the MAV primers. Of the 24 clinical samples, 3 were positive for M. avium and 6 for M. tuberculosis., Conclusions: Our results demonstrated that PCR technique could be applied for the differentiation of M. avium and M. intracellulare by specific 16S rRNA primers. In patients with advanced stage AIDS and in whom disseminated mycobacteriosis is suspected, the presence of anemia (even with negative cultures), elevated alkaline phosphatase and a median CD4 count of 15.9/mL, the diagnosis of infection by MAC should be strongly considered; we suggest that in accordance with our findings, a more precise stratification of patients in terms of their CD4 T cell counts is warranted.

Background: Community-acquired pneumonia (CAP) is one of the most common infectious causes of morbidity and mortality in children <5 years of age. The aim of the study was to clarify the bacterial etiologic diagnosis in infants with CAP., Methods: A prospective, cross-sectional and descriptive study in patients 6 months to 2 years 11 months of age with CAP with poor outcome was conducted. Patients were admitted to the Pediatric Pneumology Service and underwent bronchoscopy with bronchoalveolar lavage (BAL), taking appropriate measures during the procedure to limit the risk of contamination., Results: Aerobic bacteria isolated were Moraxella sp. 23%, Streptococcus mitis 23%, Streptococcus pneumoniae 18%, Haemophilus influenzae 12%, Streptococcus oralis 12%, and Streptococcus salivarius 12%., Conclusions: In contrast to other reports, we found Moraxella sp. to be a major bacterial pathogen, possibly because of improved detection with bronchoscopy plus BAL., (Copyright © 2015 Hospital Infantil de México Federico Gómez. Publicado por Masson Doyma México S.A. All rights reserved.)

Objective: To assess the role of polymerase chain reaction (PCR) in serum samples, in the diagnosis of osteoarticular tuberculosis (OTB) in a setting where only clinical and imaging diagnoses determine the treatment., Methods: A total of 44 consecutive serum specimens were collected from clinically suspected OTB patients, based on clinical and radiological [X-ray or magnetic resonance imaging/computed tomography] features. They were screened by in-house nested PCR. In addition, a few specimens were examined by Gram stain, acid-fast bacilli stain, histopathology and routine bacterial culture. A total of 39 specimens were collected from patients suffering from other bone diseases of nontuberculous origin and included as negative controls., Results: Of the 44 clinically suspected OTB patients, in-house nested PCR was positive in 40 (91%) cases; PCR was negative in 38 (97%) negative controls. Sensitivity and specificity of our in-house nested PCR was 90.9% and 97.4%, respectively. The PCR report was available within 48 h. It was possible to standardize serum PCR technique and in positive cases, a good correlation was observed in terms of an adequate treatment response., Conclusions: Nested PCR in serum samples is a rapid, highly sensitive and specific modality for OTB detection. PCR should be performed in addition to clinical evaluation, imaging studies, acid-fast bacilli staining, culture and histopathology diagnosis, if possible.

Background and Aims: The prevalence of infections with Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM) species in patients with immunodeficiencies in Mexico is unknown. The aim of this study was to identify, at the molecular level, the mycobacterial species most frequently affecting patients with immunodeficiencies and evaluate the genotypic diversity of MTB complex strains., Methods: We conducted a retrospective study of 97 strains in patients with the diagnosis of pulmonary (all isolates were of pathological significance) or extrapulmonary tuberculosis. PCR analysis was performed to determine whether they belonged to the MTB complex (MTC) or the Mycobacterium avium complex (MAC). Noncharacterized NTM were sequenced and, finally, MTC were genotyped by MIRUs-VNTR and spoligotyping., Results: Of the 97 mycobacterial strains isolated, 53% were M. tuberculosis, 10% M. bovis, 24% M. avium, 9% M. simiae, 2% M. kansasii and 2% M. gordonae. A great genetic diversity was found by MIRU-VNTR with the greatest polymorphism in MIRU 10, 16, 23 and 27. By spoligotyping, the predominant family was T1. Combining both methods, the association of 13 strains in four different groups was found., Conclusions: This is the first molecular analysis of mycobacteria isolated from patients with immunodeficiencies in Mexico, describing the prevalence of different mycobacterial species in this population. A great genetic diversity of MTB strains was identified. This is also the first report in Mexico describing clinically important isolates of M. simiae., (Copyright © 2013 IMSS. Published by Elsevier Inc. All rights reserved.)

Objective: to determine the prevalence of opportunistic microorganisms and microbial flora in neutropenic enterocolitis in oncohematological pediatric patients., Methods: a prospective and observational study was done. Patients with diagnosis of acute leukemia and neutropenia were included. Stool cultures were taken to identify microorganisms and microbial flora. A χ(2) test with Yates corrections and Fisher exact test were used in the statistical analysis., Results: 21 patients were included (12 male, 57.1 %). The stool cultures showed that 68 % of microorganisms were Gram-negative. The presence of microorganisms Gram-positive was 20 %, 6 % for Candida sp.; 3 % for Cryptosporidium sp.; and in 3 % were acid fast bacilli. Staphylococcus epidermidis, Enterobacter sp., and Escherichia coli were presented in pure culture. No association was found between Gram-positive and Gram-negative microorganisms with age, white cell count or pure or mixed cultures., Conclusions: although Gram-negative microorganisms were the most frequent, Gram-positive and other microorganisms that are not detected habitually in feces culture were isolated.

Objective: To determine the polymorphisms of Interleukin-10 (IL-10) (-592, -1082) in pulmonary tuberculosis (PTB) with and without type 2 diabetes (T2D)., Methods: We studied a Mexican mestizo population of 37 patients with TB in remission (TBr) and 40 with active pulmonary TB (PTB), 21 patients with TB + T2D, 47 blood donors accepted, and 13 healthy health-care workers with tuberculin skin test positive. Determination of IL-10 polymorphisms was performed by real-time Polymerase chain reaction., Results: IL-10-592C/A presented in a greater proportion in healthy individuals than in patients with type 2 diabetes and TB in a not quite significant statistically manner. IL-10-1082A/A presented more frequently in the group of patients with both diseases, not being statistically significant in comparison with the group of healthy subjects., Conclusions: This study describes two important new findings. First, it reveals that the IL-10 (-592 A/A and -592 C/C) polymorphisms were found in a greater proportion in a group of patients with T2D and TB than in healthy subjects. Second, the study provides evidence that the (-1082 G/G) polymorphism presented with greater frequency in healthy individuals than in patients with both of these diseases., (Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.)

Objective: To assess the usefulness of IGRA test (QuantiFERON(®)-Cell mediated immune) compared with the tuberculin skin test., Methods: A cross-sectional study was carried out in Mexico, 25 infected patients with HIV-AIDS and the suspicion or with latent tuberculous infection (LTBI) who were >18 years of age and without treatment for tuberculosis (TB), were enrolled in the study., Results: Median cluster of differentiation (CD4) count was 364 cells/μ L and median HIV viral load was 50 copies/mL. Overall, 20 patients (80%) had at least one positive diagnostic test for LTBI: four (16%) had a positive tuberculin skin test and 19 (76%), a positive QuantiFERON(®)-tuberculosis., Conclusions: No agreement is found between the two diagnostic tests: k = -0.004, 95% confidence interval (-0.2219, 0.2210). Additional longitudinal studies among HIV-infected populations with high prevalence of TB are needed to further assess the usefulness of IGRAs in this patient population., (Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.)

Objective: To investigate the frequency of the single-base change polymorphic variants identified in tumor necrosis factor (TNF) gene (-308 G/A) and lymphotoxin alpha (LTA) (+252 G/A) in patients with type 2 diabetes (T2D)., Methods: A prospective study in a Mexican-mestizo population of 51 patients with T2D and 48 healthy subjects was carried out. We took a peripheral blood sample from each individual for identification of the polymorphic genotypes by polymerase chain reaction., Results: The genotype distribution in T2D was: TNF alpha homozygous 0%; TNFG/A heterozygous 20%; TNFG homozygous 80%., Conclusions: In regards to the TNF -308 G/A genotype, we found a significant difference (p = 0.012) with a bigger frequency in the group of patients. The health controls showed a higher frequency of TNF -308 G/G genotype (p = 0.034).

Objective: to determine the relation between IL6, IL10 and TNFa serum levels in a cohort of patients with type 2 diabetes (T2D) and severe soft tissue infections (STI), with severity and mortality factors., Methods: A. comparative and transversal, study with 15 adult patients, any gender, with T2D and STI were done. A T2D control group of 20 patients without STI was included. Apache II Score, glycemia and by ELISA, IL6, IL10 and TNFa, were determined., Results: in all patients, it was a correlation at beginning between glycemia and IL6 (r = 0.67, IC 95 % 0.24-0.88), as soon as glycemia and Apache II, (r = 0.59, IC 95 % 0.11-0.83)., Conclusions: although IL6 was very usefulness, it is not a routine test in clinical laboratory and it is expensive, but in medical practice, it could be possible to evaluate these patients with Apache II Score and glycemia. However, in STI, the values of IL6 and IL10 were highly significant. It is likely that IL6 is a marker of poor outcome.

Objective: In this study are evaluated the usefulness of the buffy coat smear and panbacterial polymerase chain reaction (PCR) as diagnostic tests in the early detection of neonatal sepsis., Material and Methods: It was studied 49 patients aged up to 28 days who were hospitalized in the Intensive Care Unit (ICUs) of the Neonatology, with a clinical diagnosis of neonatal sepsis and 49 umbilical cord samples of healthy newborns. Blood cultures and 50 microL of plasma were taken for the DNA and performance of the broad-range PCR primer system (panbacterial PCR). Simultaneously, were taken three capillaries with blood for the leukocyte layer (buffy coat) smear, we performed three stains: Gram; Löeffler blue methylene (LBM), and acridine orange (AO). Statistical analysis included sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) against the clinical diagnosis., Results: With respect to stains of buffy coat smear, they resulted very specific, from 90-97%, with 64-75% sensitivity, 87-94% PPV, and 77-82% NPV. In inverse fashion, PCR resulted very sensitive at 96%, with 91% specificity, 92% PPV, and 96% NPV., Conclusions: Buffy coat smear stains are easy, fast, and specific, while that of PCR was highly sensitive. Thus, both can be utilized as diagnostic tests.

Background: The phagocytic function in pulmonary tuberculosis (PTB) and Type 2 diabetes (T2D) has been explored mainly in macrophages but not in polymorphonuclears (PMN). The purpose of this study was to determine the functional status of PMN leukocytes in patients with pulmonary tuberculosis (PTB), Type 2 diabetes (T2D), and in patients with both diseases., Methods: An observational, prospective, and comparative study was carried out. 30 ambulatory patients with T2D, 10 with PTB undergoing treatment and 10 patients with PTB and T2D, and 44 healthy subjects were studied. PMN leukocytes were separated, the capacity of these cells to produce hydrogen peroxide and to reduce nitroblue tetrazolium (NBT) in response to stimulus with the phorbolic ester of myristic acid (PMA) was measured; and the capacity of PMN leukocytes to adhere to surfaces was determined., Results: Concerning the test for adherence, on comparing healthy subjects with patients with T2D+PTB, we observed a clear decrease in cellular adherence in the group of patients with both diseases; it was statistically significant (p = 0.007).With regard to phagocytic function, we observed that in NBT reduction as well as in hydrogen peroxide production, statistically significant differences were not obtained on comparing healthy subjects with any of the three groups of patients., Conclusions: We observed a clear decrease in cellular adherence when both diseases co-exist. These results could indicate the need for the co-existence of T2D and TB to cause deterioration in the cells' adherence activity. The microtechniques employed permit the evaluation in a practical manner of certain phagocytic-activity expressions.

Objective: We standardized the RT-PCR panviral CSF and determined its applicability in detecting acute enterovirus infection in the central nervous system in children under 15 years., Material and Methods: RT-PCR was performed directly in CSF samples of 10 pediatric patients with suspected CNS infection and 9, with different conditions of the central nervous system., Results: 80% (8/10) of RT-PCR samples were positive for enterovirus in patients with suspected CNS infection and no sample was positive in patients with different ailments., Conclusions: Since enteroviruses are among the main etiologies of pediatric encephalitis, RT-PCR could be particularly useful for rapid detection in CSF.

Background and Aims: The Mexican population has a distinct capacity for the expression of tumor necrosis factor (TNF), a cytokine that plays a cardinal role in Kawasaki disease (KD), particularly in those who develop coronary aneurysms. It is important to identify, in Mexican pediatric patients, the association of the frequency of TNF. This study determined the association of TNF -308 and lymphotoxin-alpha (LTA) +252 polymorphisms in Mexican pediatric patients with KD and coronary aneurysms (CA)., Methods: We conducted a cross-sectional, analytical study in 48 children with KD, 22 with CA. Control samples were obtained from 61 aged-matched children. We took a peripheral blood sample and extracted genomic DNA from all children participating in the study. Using restriction factor length polymorphism-polymerase chain reaction (RFLP-PCR), we performed determination of TNF -308 and LTA +252., Results: There was no difference in frequency between the study groups for genotype LTA +252 (OR 0.37, 95% CI, 0.06-2, p = 0.44) or between groups for KD with or without coronary aneurysms for both polymorphisms. In subjects with KD, we did not observe the heterozygous genotype of TNF -308, the difference being significant (OR 12, 95% CI, 4.8-30.4, p = 0.0001) using the χ(2) test with the continuity correction on comparison with the control group., Conclusions: Comparative analysis by group did not show a significant difference in the frequency of the alleles and genotypes between KD with CA vs. KD without CA vs. controls, for both TNF -308 and LTA +252., (Copyright © 2011 IMSS. Published by Elsevier Inc. All rights reserved.)

This study compared the frequency of the genetic polymorphisms of tumor necrosis factor (TNF) in pulmonary tuberculosis without and with response to treatment. We carried out an observational, prospective, comparative study. Three groups were studied: healthy subjects, responders, and non-responders to directly observed treatment short-course. We took a peripheral blood sample for identification of polymorphic genotypes TNF -308G/A and lymphotoxin A (LTA) +252G/A by polymerase chain reaction, and their later digestion with the Nco1 restriction enzyme. We studied a total of 138 subjects: 42 (non-responders) and 48 in each of the remaining groups. Healthy subjects had significantly high frequency of the LTA +252A allele compared to groups of patients and could be related with protection from the disease. Patients had higher frequency of the non-polymorphic allele LTA +252G than healthy subjects. With regard to LTA +252G/A genotype, we did find a significant difference with a greater frequency in the group of patients. The LTA +252G/A genotype was associated with impaired response to treatment.

Atherosclerosis is a multifactor disease. Lately, infectious factors such as C. pneumoniae have been found to be involved. To determine whether the infection by C. pneumoniae is a risk factor for atherosclerosis in patients with AIDS. Case-control study on 43 patients with AIDS under HAART (16 cases and 27 controls). To document atherosclerosis, a carotid and transcranial Doppler ultrasound was performed. Anti-C pneumoniae antibodies were searched using a microimmunofluorescence test for IgM and IgG levels. To study the associations with risk of atherosclerosis, Odds Ratios were calculated for each IgG anti-C. pneumoniae antibody titre. A titre of 1:64 significantly increased the risk of atherosclerosis. These results suggest that hypertriglyceridemia and C. pneumoniae infection coexistence significantly increases the risk of atherosclerosis. The inverse geometric average of the antibodies titre against C. pneumoniae in individuals with atheromatous plaque fell to 64, two titres above the controls. This difference turned out to be statistically significant. Exposure to C. pneumoniae with antibodies (IgG) should be considered in any HIV diagnosed patient as a risk factor for atherosclerosis, having found that the inverse geometric averages of antibodies titre are significantly different comparing cases and controls, especially in patients with dyslipidemia, hypertriglyceridemia or in patients whose treatments could cause these conditions. In patients with concomitant hypertriglyceridemia, the association increases up to three times. It is advisable that AIDS patients take a serological test to determine exposure to C. pneumoniae, and to assess treatment options.

Objective: To assess the performance in the clinical setting of the MB/BacT system for isolation of Mycobacterium tuberculosis and to verify by PCR., Material and Methods: The study included 272 sputum samples from 208 patients with the presumptive diagnosis of pulmonary tuberculosis. ZN was made, culture in Löwenstein-Jensen medium, MB/BacT and PCR., Results: Thirty-nine samples were positive by culture in Löwenstein-Jensen, and 42 using the MB/BacT system. Positive cultures in the MB/BacT system were verified by acid-fast bacilli staining and PCR. Mycobacterial identification in the MB/BacT took 8 to 46 days (mean 16 days), while the Löwenstein-Jensen culture ranged between 21 and 63 days (mean 35 days). These results show that the MB/BacT semiautomated system is reliable and faster than the manual culture method and can be used as an alternative for the primary identification of Mycobacterium tuberculosis. The PCR assay allows the fast and exact identification of Mycobacterium tuberculosis directly from positive liquid medium.

Background: Antimyocardium antibodies (AMA) have been observed in patients with acute myocardial infarction (AMI). At present it is unknown if these AMA play a role in the pathogenesis of acute or chronic myocardial damage or if they are only produced as a result of tissue destruction, disappearing later without clinical manifestations. However, some studies have shown that patients with AMI and AMA have higher possibilities of presenting heart failure and death. The aim of the present study was to determine if there is an association (odds ratio, OR) between AMA and death in patients with AMI., Methods: The design was a cross-sectional study. One hundred patients with acute myocardial infarction were recruited consecutively at the Coronary Care Unit of the Hospital de Especialidades at La Raza National Medical Center, IMSS, Mexico City. IgG antimyocardium antibodies were identified using the enzyme-linked immunosorbent assay technique (ELISA)., Results: Of the 100 patients studied, 81 were males and 19 females, with an average age of 58 +/- 11 years. Of the population studied, 44% showed AMA. It was observed that patients with positive AMA had an OR for heart failure of 3.40 (CI 95% 0.97-12.5, p = 0.06) and for death of 7.94 (CI 95%, 1.49-56.1, p = 0.003). This variable was analyzed with other confounding variables using logistic regression, and an OR of 11.8 (CI 95% 1.63-86.3, p = 0.001) was obtained., Conclusions: AMAs were an independent predictive factor for mortality in patients with AMI.

Objective: The primary aim of this study was to determine whether antibodies against Chlamydophila pneumoniae in patients with acute myocardial infarction (AMI) and coronary risk factors are associated with death., Material and Methods: A cross-sectional study was conducted among 100 patients hospitalized in the Coronary Unit of Centro Medico La Raza Hospital of the Mexican Institute of Social Security, between 1999 and 2000. Subjects were males and females older than 18 years, diagnosed with AMI and coronary risk. Antibodies against Chlamydophila pneumoniae, Chlamydophila psitacii and Chlamydia trachomatis were measured using an indirect microinmunofluorescence assay. In addition, blood samples from 33 patients from the original group were taken when the patients were discharged from the hospital,and 3 months after their myocardial infarction. Data analysis consisted of geometric means and standard deviations as well as odds ratios with 95% confidence intervals., Results: Seventy percent of patients presented antibodies against Chlamydophila pneumoniae. Antibodies against Chlamydophila psittaci and Chlamydia trachomatis were not identified. No statistically significant association was found between antibodies and death in these patients with coronary risk factors and AMI. In the subgroup of 33 individuals 25 had antibodies against Chlamydophila pneumoniae and in 83% of them antibodies decreased three months after the AMI event., Conclusions: Even though patients with coronary risk factors and AMI had an increased seropositivity for Chlamydophila pneumoniae it was not significantly associated with death.