Effect of the buccal fat pad in the prevention of zygomatic implant surgery postoperative complications: A pilot study.

BACKGROUND: Zygomatic implants have been used to treat severe atrophy maxilla. Since its description, the technique has been improved in order to reduce patient morbidity as well as prosthesis rehabilitation time. Despite the improvements in the procedure, zygomatic implant treatments still have complications related to the peri-implant soft-tissue; a probing depth greater than 6 millimeter (mm) and a prevalence of bleeding on probing of 45% have been described. The mobilization of the buccal fat has been used to manage different oral and maxillofacial soft-tissue pathologies. The aim of this study was to assess whether the buccal fat pad might prevent mucosal dehiscence and avoid potential postoperative complications when is placed covering the body part of the zygomatic implants. MATERIAL AND METHODS: In this pilot study, 7 patients were enrolled and a total of 28 zygomatic implants were placed and evaluated during a 12-month follow-up period. Surgical sites were randomly divided into two groups before implant placement: control group (A; in which no buccal fat pad was applied) and experimental group (B). Peri-implant soft tissue thickness difference, pain using a Visual Analog Scale (VAS), swelling, hematoma, buccal soft tissue healing and sinusitis, were evaluated. The implant survival rate was determined according the Aparicio success criteria and compared between the control and experimental procedure. RESULTS: A nonstatistical difference was found between groups regarding to pain. The experimental group showed higher soft-tissues thickness (p= 0.03) and the implant survival rate was 100% in both groups. CONCLUSIONS: The mobilization of the buccal fat pad to cover the body of the zygomatic implants increases peri-implant soft-tissue thickness, without increasing the postoperative pain.

Evaluation of GenoFlow DR-MTB Array Test for Detection of Rifampin and Isoniazid Resistance in Mycobacterium tuberculosis.

The aim of this study was to evaluate the GenoFlow DR-MTB array test (DiagCor Bioscience, Hong Kong) on 70 cultured isolates and 50 sputum specimens. The GenoFlow array test showed good sensitivity and specificity compared to the phenotypic Bactec 460TB. This array accurately detected mutations inrpoB,katG, andinhAassociated with resistance to rifampin and isoniazid.

GenoType MTBDRsl for molecular detection of second-line-drug and ethambutol resistance in Mycobacterium tuberculosis strains and clinical samples.

The purpose of this study was to evaluate the GenoType MTBDRsl assay (Hain Lifescience GmbH, Nehren, Germany) for its ability to detect resistance to fluoroquinolones (FLQ), injectable second-line antibiotics [kanamycin (KM) and capreomycin (CM)], and ethambutol (EMB) in Mycobacterium tuberculosis clinical strains and directly in clinical samples. A total of 34 clinical strains were characterized with the Bactec 460 TB system. Fifty-four clinical samples from 16 patients (5 were smear negative and 49 were smear positive) were also tested directly. The corresponding isolates of the clinical specimens were also analyzed with the Bactec 460TB. When there was a discrepancy between assays, pyrosequencing was performed. The overall rates of concordance of the MTBDRsl and the Bactec 460TB for the detection of FLQ, KM/CM, and EMB susceptibility in clinical strains were 72.4% (21/29), 88.8% (24/27), and 67.6% (23/34), whereas for clinical samples, rates were 86.5% (45/52), 92.3% (48/52), and 56% (28/50), respectively. In conclusion, the GenoType MTBDRsl assay may be a useful tool for making early decisions regarding KM/CM susceptibility and to a lesser extent regarding FLQ and EMB susceptibility. The test is able to detect mutations in both clinical strains and samples with a short turnaround time. However, for correct management of patients with extensively drug-resistant tuberculosis, results must be confirmed by a phenotypical method.

Pyrosequencing for rapid molecular detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis strains and clinical specimens.

The aim of this study was to evaluate a pyrosequencing method for the detection of Mycobacterium tuberculosis isolates resistant to rifampin and isoniazid using both clinical strains and clinical samples, comparing the results with those of the Bactec 460TB and GenoType MTBDRplus assays. In comparison to Bactec 460TB as the gold standard, the sensitivity of pyrosequencing for detecting isoniazid and rifampin resistance was 76.9% and 97.2%, respectively, for clinical strains, and the specificity was 97.2 and 97.9%, respectively. For clinical specimens, the sensitivity and specificity for both drugs were 85.7% and 100%, respectively. The overall concordance between pyrosequencing and the GenoType MTBDRplus assay for clinical strains was 99.1%, and for clinical samples, it was 98.2%. Pyrosequencing is a valuable tool for rifampin and isoniazid resistance detection.

Characterization of Mycobacterium tuberculosis Beijing isolates from the Mediterranean area.

BACKGROUND: The Beijing lineage of Mycobacterium tuberculosis is causing concern due to its global distribution and its involvement in severe outbreaks. Studies focused on this lineage are mainly restricted to geographical settings where its prevalence is high, whereas those in other areas are scarce. In this study, we analyze Beijing isolates in the Mediterranean area, where this lineage is not prevalent and is mainly associated with immigrant cases. RESULTS: Only 1% (N = 26) of the isolates from two population-based studies in Spain corresponded to Beijing strains, most of which were pan-susceptible and from Peruvian and Ecuadorian patients. Restriction fragment length polymorphism typing with the insertion sequence IS6110 identified three small clusters (2-3 cases). Mycobacterial interspersed repetitive unit-variable number tandem repeat typing (MIRU-15) offered low discriminatory power, requiring the introduction of five additional loci. A selection of the Beijing isolates identified in the Spanish sample, together with a sample of Beijing strains from Italy, to broaden the analysis context in the Mediterranean area, were assayed in an infection model with THP-1 cells. A wide range of intracellular growth rates was observed with only two isolates showing an increased intracellular replication, in both cases associated with contained production of TNF-alpha. No correlation was observed between virulence and the Beijing phylogenetic group, clustered/orphan status, or resistance. The Beijing strain responsible for extensive spread on Gran Canaria Island was also identified in Madrid, but did not lead to secondary cases and did not show high infectivity in the infection model. CONCLUSIONS: The Beijing lineage in our area is a non-homogeneous family, with only certain highly virulent representatives. The specific characterization of Beijing isolates in different settings could help us to accurately identify the virulent representatives before making general assumptions about this lineage.

Multidrug-resistant tuberculosis in a low-incidence region shows a high rate of transmission.

OBJECTIVE: To describe the characteristics of patients with multidrug-resistant tuberculosis (MDR-TB), a descriptive prospective study was carried out applying a combination of exhaustive conventional epidemiology with molecular genotyping. SETTING: All patients diagnosed with MDR-TB in Galicia, Spain, between 1998 and 2004 were included in the study. DESIGN: Of 9895 diagnosed cases of TB, 58 were MDR-TB (0.59%). The site of disease was pulmonary in 56 cases and 46 were smear-positive. Only two cases were co-infected with the human immunodeficiency virus (HIV) and seven were immigrants. Twenty-five (43%) had received previous TB treatment. These cases presented more risk factors for treatment default and a lower frequency of contact with cases of MDR-TB. RESULTS: Genotyping analysis was performed in 57 patients, showing evidence of four clusters (30 patients, 52.6%), each with identical genetic patterns. The patients included in the clusters were younger, and most had primary forms or had had contact with another case of MDR-TB, especially in hospital. Neither the Beijing/W nor the B strain was identified. CONCLUSION: There is a low prevalence of MDR-TB in Galicia. Unlike previous studies, there was a high rate of transmissibility, including nosocomial transmission. Transmission is not associated with HIV or previously reported strains with a high capacity for transmission.

Genotyping of Mycobacterium tuberculosis over two periods: a changing scenario for tuberculosis transmission.

OBJECTIVE: To evaluate changes in the molecular epidemiology of tuberculosis (TB) in Zaragoza, Spain, over a decade that has seen large social and health changes, including the attenuation of the human immunodeficiency virus (HIV) epidemic and increased immigration. DESIGN: A population-based molecular study was conducted using standard restriction fragment length polymorphism IS6110 typing that included all patients with bacteriologically confirmed TB living in the Zaragoza area from 2001 to 2004. The current situation was compared with that described in a previous study from 1993 to 1995. RESULTS: A total of 454 Mycobacterium tuberculosis isolates were genotyped; 239 (52.6%) were grouped in 45 clusters composed of 2 to 85 isolates. Independent risk factors for clustering were identified. The main differences with the previous study were the increase of TB cases among immigrants, a decrease in HIV-TB co-infected patients and the occurrence of a large TB outbreak involving 85 patients (M. tuberculosis Zaragoza [MTZ] strain). CONCLUSION: A change in the epidemiological pattern of TB has been observed in the last years. TB transmission is more common among the Spanish-born population, while foreign birth is significantly less associated with clustering. A single epidemic strain caused 18.7% of all TB cases.

PyroTyping, a novel pyrosequencing-based assay for Mycobacterium tuberculosis genotyping.

We developed a novel method, PyroTyping, for discrimination of Mycobacterium tuberculosis isolates combining pyrosequencing and IS6110 polymorphism. A total of 100 isolates were analysed with IS6110-restriction fragment length polymorphism (RFLP), spoligotyping, mycobacterial interspersed repetitive units - variable number tandem repeats (MIRU-VNTR), and PyroTyping. PyroTyping results regarding clustering or discrimination of the isolates were highly concordant with the other typing methods performed. PyroTyping is more rapid than RFLP and presents the same discriminatory power, thus, it may be useful for taking timely decisions for tuberculosis control.

Peritoneal tuberculosis due to Mycobacterium caprae.

The incidence of tuberculosis in humans due to Mycobacterium caprae is very low and is almost confined to Europe. We report a case of a previously healthy 41-year-old Moroccan with a 6 month history of abdominal pain, weight loss, fatigue and diarrhea. A diagnosis of peritoneal tuberculosis due to M. caprae was made.

Mycobacterial diversity causing multi- and extensively drug-resistant tuberculosis in Djibouti, Horn of Africa.

On detecting a high prevalence of multidrug-resistant tuberculosis (TB) in Djibouti, 32 Mycobacterium tuberculosis isolates of patients hospitalised in the TB referral centre of the capital were genotyped. A high variety of M. tuberculosis lineages, including lineage 1, Indo-Oceanic, lineage 2, East-Asian, lineage 3, East-African Indian and lineage 4, Euro-American, were detected.

Ten-year population-based molecular epidemiological study of tuberculosis transmission in the metropolitan area of Madrid, Spain.

SETTING: The epidemiology of tuberculosis (TB) in urban populations is changing. Combining conventional epidemiological techniques with DNA fingerprinting of Mycobacterium tuberculosis can improve our understanding of how TB is transmitted. OBJECTIVE: To improve the definition of molecular epidemiology of TB over 10 years in an area of Europe not previously studied. DESIGN: A population-based retrospective study was conducted in the Autonomous Community of Madrid, Spain, from 1992 to 1998; from 1999 to 2001, the study was prospective. The study population consisted of all patients for whom positive culture and full clinical and demographic data were available. All strains were typed by RFLP. Non-clustered patients were compared with clustered patients and studied using univariate analysis and a logistic regression model. RESULTS: Of 448 patients studied, 228 (50.7%) were clustered. Youth was the strongest risk factor associated with clustering. Pleural effusion was also found to be associated with clustering. An epidemiological link was found in only 85 (37.4%) of the 228 patients belonging to a cluster. CONCLUSION: Youth and pleural effusion were identified as risk factors for clustering. These findings may help adjust TB control and contact tracing strategies.

AID TB resistance line probe assay for rapid detection of resistant Mycobacterium tuberculosis in clinical samples.

OBJECTIVES: To determine the sensitivity and specificity of AID TB Resistance line probe assay (AID Diagnostika, Germany) to detect Mycobacterium tuberculosis and its resistance to first- and second-line drugs in clinical samples using BACTEC 460TB as the reference standard. METHODS: The test consists on three strips to detect resistance to isoniazid/rifampicin, fluoroquinolones/ethambutol, and kanamycin/amikacin/capreomycin/streptomycin, respectively. This test was performed on 65 retrospectively selected clinical samples corresponding to 32 patients. RESULTS: A valid result was obtained for 92.3% (60/65), 90.8% (59/65) and 78.5% (51/65) of the samples tested, considering the three strips, respectively. Global concordance rates between AID and BACTEC for detecting resistance to isoniazid, rifampicin, fluoroquinolones, ethambutol, kanamycin/capreomycin and streptomycin were 98.3% (59/60), 100% (60/60), 91.5% (54/59), 72.9% (43/59), 100% (51/51) and 98.0% (50/51), respectively. Regarding the discordant results obtained between AID and BACTEC, the alternative molecular methods performed (GenoType MTBDRplus, GenoType MTBDRsl [Hain Lifescience, Germany] and/or pyrosequencing) confirmed the genotypic result in 90.9% (20/22) of the cases. CONCLUSIONS: AID line probe assay is a useful tool for the rapid detection of drug resistance in clinical samples enabling an initial therapeutic approach. Nevertheless, for a correct management of drug resistant tuberculosis patients, molecular results should be confirmed by a phenotypic method.

Diagnostic accuracy study of multiplex PCR for detecting tuberculosis drug resistance.

OBJECTIVE: To study the diagnostic accuracy of a multiplex real-time PCR (Anyplex II MTB/MDR/XDR, Seegene, Corea) that detects Mycobacterium tuberculosis resistant to isoniazid (INH), rifampicin (RIF), fluoroquinolones (FLQ) and injectable drugs (kanamycin [KAN], amikacin [AMK] and capreomycin [CAP]) in isolates and specimens. METHODS: One hundred fourteen cultured isolates and 73 sputum specimens were retrospectively selected. Results obtained with multiplex PCR were compared with those obtained with BACTEC. Discordant results between multiplex PCR and BACTEC were tested by alternative molecular methods. RESULTS: Sensitivity and specificity of multiplex PCR for detecting drug resistance in isolates were 76.5% and 100%, respectively, for INH; 97.2% and 96.0%, respectively, for RIF; 70.4% and 87.9%, respectively, for FLQ; 81.5% and 84.8%, respectively, for KAN; 100% and 60%, respectively, for AMK, and 100% and 72.3%, respectively, for CAP. Sensitivity and specificity of Anyplex for detecting drug resistance in specimens were 93.3% and 100%, respectively, for INH; 100% and 100%, respectively, for RIF; 50.0% and 100%, respectively, for FLQ; and 100% and 94.4%, respectively, for both KAN and CAP. Among the discordant results, 87.7% (71/81) of results obtained with the multiplex PCR were concordant with at least one of the alternative molecular methods. CONCLUSIONS: This multiplex PCR may be a useful tool for the rapid identification of drug resistant tuberculosis in isolates and specimens, thus allowing an initial therapeutic approach. Nevertheless, for a correct management of patients, results should be confirmed by a phenotypic method.

Transmission between HIV-infected patients of multidrug-resistant tuberculosis caused by Mycobacterium bovis.

OBJECTIVE: To investigate outbreaks of multidrug-resistant tuberculosis (TB) by using DNA fingerprint databases. DESIGN: Investigation of two outbreaks of multidrug-resistant TB in separate hospitals in Spain by restriction fragment length polymorphism (RFLP) and spoligotyping. Outbreak strains were compared with more than 1500 RFLPs of Mycobacterium tuberculosis complex strains isolated in Spain and 6000 RFLPs from 30 different countries. METHODS: Standardized IS6110 DNA fingerprinting and 'spoligotyping' was used to type multidrug-resistant isolates belonging to the M. tuberculosis complex amongst the outbreak cases. The DNA types were matched against DNA fingerprint databases in Spain and The Netherlands. RESULTS: The DNA typing analysis indicated that a single multidrug-resistant Mycobacterium bovis strain was responsible for a nosocomial outbreak in a hospital in Spain involving at least 16 HIV-infected patients with non-treatable to multidrug-resistant TB. Introduction of the fingerprint type of this strain to the international database revealed a single matching strain. This strain was also isolated from an HIV-infected patient in The Netherlands who had died from multidrug-resistant TB. This patient had previously been hospitalized in Spain, where a multidrug-resistant TB nosocomial outbreak involving 20 HIV-infected patients was ongoing. The strains causing this outbreak were also identified as M. bovis with an identical DNA pattern to those strains isolated in the Spanish hospital and the patient in The Netherlands. CONCLUSIONS: The use of centralized DNA databases can help to identify rapidly the origin and transmission routes of multidrug-resistant TB across international boundaries and the potential use of such an early warning surveillance system for investigation of nosocomial multidrug-resistant TB outbreaks between HIV-infected patients. To our knowledge this is the first report of transmission of multidrug-resistant M. bovis between hospitals.

Prolonged survival of an HIV-infected patient with multidrug-resistant Mycobacterium bovis infection treated with surgical resection.

We describe a case of multidrug-resistant tuberculosis due to Mycobacterium bovis in a human immunodeficiency virus-infected woman with good immunologic status. The patient presented with a hard mass measuring 10 cm in diameter on the lower left ribs and a lung nodule measuring 3 cm in diameter in the left superior lobe. No adequate pharmacological treatment was available. Both lesions were surgically resected. The patient has remained asymptomatic (without fever, cough, lymphadenopathy, or cutaneous masses) for 20 months, after discharge from the hospital.

Molecular epidemiology of tuberculosis in Elche, Spain: a 7-year study.

The epidemiology of tuberculosis in Elche (Spain) was studied by restriction fragment-length polymorphism (RFLP) typing of clinical isolates of Mycobacterium tuberculosis over a 7-year period. A total of 165 isolates was typed and the clinico-epidemiological data of the patients were studied retrospectively. An overall cluster aggregation of 52.4% was found, rising to 71.43% in HIV-positive patients. There was greater aggregation in younger patients, but no statistically significant differences when other variables were analysed. The percentage of aggregation was higher than in other studies in Spain and this may be due to the longer time period of this study. The high percentage of aggregation in young patients and in thosewho were HIV-positive suggests increased recent transmission in both groups.

The molecular epidemiology of tuberculosis in Zaragoza, Spain: a retrospective epidemiological study in 1993.

SETTING: The incidence of tuberculosis (TB) in Spain is one of the highest in Europe. In Zaragoza region the incidence rate of tuberculosis and the acquired immune deficiency syndrome (AIDS) are close to the national average. OBJECTIVE: To better define the molecular epidemiology of tuberculosis in an area of Europe where this has not been previously studied. DESIGN: A retrospective epidemiological study on tuberculosis was conducted in Zaragoza, a region of Spain, in 1993. The study population consisted of 226 patients from whom positive culture and complete clinical and demographic data were available. Mycobacterium tuberculosis strains were typed by standard restriction fragment length polymorphism (RFLP). A cluster was defined as two or more isolates with identical RFLP patterns when five or more copies of IS6110 are present. The 137 non-clustered patients were compared with the 89 clustered patients and studied by using univariate analysis. RESULTS: Thirty-nine percent of the patients were clustered, suggesting possible recent transmission. Infection with drug-resistant M. tuberculosis was associated with a decreased risk of being in a cluster. The strains isolated from human immunodeficiency virus (HIV)-positive patients were not associated with clustering. We found that immigration was not a major determinant in the total number of TB cases. CONCLUSION: Immigration, HIV and drug resistance were not associated with recent transmission. More than 50% of the clusters contained two or three patients, indicating that small outbreaks were responsible for most of the tuberculosis cases. Our RFLP typing results indicate that a TB control programme should be implemented in Spain in order to lower transmission of TB.

The Spanish multidrug resistant tuberculosis network.

The network to monitor the spread of multidrug resistance tuberculosis (MDR-TB) in Spain based on genomic typing and set up in January 1998 benefits from the participation of about 90% of the laboratories of the national health system. Of the 94 MDR-TB pa

Epidemiology of subclinical salmonellosis in wild birds from an area of high prevalence of pig salmonellosis: phenotypic and genetic profiles of Salmonella isolates.

The epidemiology of subclinical salmonellosis in wild birds in a region of high Salmonella prevalence in pigs was studied. Three hundred and seventy-nine faecal samples from 921 birds trapped in 31 locations nearby pig premises, and 431 samples from 581 birds of 10 natural settings far from pig farms were analysed for the presence of Salmonella spp. Positive samples were serotyped and analysed for antimicrobial resistance (AR). Phage typing and pulsed-field gel electrophoresis (PFGE) on Salmonella Typhimurium isolates were also carried out. The overall proportion of Salmonella-positive samples was 1.85% (95% CI=0.93, 2.77). Salmonella isolation was positively associated with samples collected from birds in the proximity of a pig operation (OR=16.5; 95% CI=5.17, 52.65), and from non-migratory (or short-distance migration) birds (OR=7.6; 95% CI=1.20, 48.04) and negatively related to mostly granivorous birds (OR=0.4; 95% CI=0.15, 1.13). Salmonella Typhimurium was the most prevalent serotype and four different XbaI PFGE patterns were observed that matched the four phage types identified (U310, U311, DT164 and DT56). Only 20% of the strains showed multi-AR. In three farms, a high degree of homogeneity among isolates from different birds was observed. These findings suggested that pig farms may act as amplifiers of this infection among wild birds, and the degree of bird density may have much to do on this transmission. Some of the Salmonella serotypes isolated from bird faeces were of potential zoonotic transmission and associated with AR. Monitoring salmonellosis in wild bird is advised.