groEL is a suitable genetic marker for detecting Vibrio parahaemolyticus by loop-mediated isothermal amplification assay.

A groEL gene-based loop-mediated isothermal amplification (LAMP) assay was developed to detect Vibrio parahaemolyticus in contaminated seafood and water. The assay was optimized and conducted at 63°C for 40 min using Bacillus stearothermophilus (Bst) DNA polymerase, large fragment. Amplification was analysed via multiple detection methods, including opacity, formation of white precipitate, DNA intercalating dyes (ethidium bromide and SYBR Green I), metal ion-binding indicator dye, calcein, and 2% agarose gel electrophoresis. A characteristic ladder-like band pattern on agarose gel and the desired colour changes when using different dyes were observed in positive cases, and these were species-specific for V. parahaemolyticus when compared with other closely related Vibrio spp. The limit of detection (LoD) of this assay was 100 fg per reaction, 100-fold higher than that for conventional polymerase chain reaction (PCR). When tested on artificially contaminated seafood and seawater, the LoDs of the LAMP assay were 120 and 150 fg per reaction respectively, and those of conventional PCR were 120 and 150 pg per reaction respectively. Based on our results, the groEL gene-based LAMP assay is rapid, specific, sensitive, and reliable for detecting V. parahaemolyticus, and it could be used in field diagnosis. SIGNIFICANCE AND IMPACT OF THE STUDY: The loop-mediated isothermal amplification (LAMP) assay using groEL gene (an abundant, highly conserved gene and member of the groESL chaperone gene family) provided rapid, species-specific and highly sensitive method for detecting Vibrio parahaemolyticus, the leading causal agent of seafood-borne diseases worldwide. Moreover, groEL LAMP revealed high efficiency than conventional PCR assay for V. parahaemolyticus using template both from pure culture and artificially contaminated seafood and water, which indicated the applicability in the field and environmental screening purpose for the organism.

Complete Genome Sequence of Mycobacterium tuberculosis Clinical Isolate Spoligotype SIT745/EAI1-MYS.

Mycobacterium tuberculosis is known to cause pulmonary and extrapulmonary tuberculosis. This organism showed special phylogeographical specificity. Here, we report the complete genome sequence of M. tuberculosis clinical isolate spoligotype SIT745/EAI1-MYS, which was isolated from a Malaysian tuberculosis patient.

Modulating the effects of diabetes on osseointegration with aminoguanidine and doxycycline.

BACKGROUND: The current knowledge of wound healing around implant surfaces is quite limited, particularly as it relates to the effects of systemic diseases such as diabetes. The purpose of our research is to histologically evaluate the effects of aminoguanidine and doxycycline in the modification of peri-implant wound healing around endosseous implants in diabetic rats. METHODS: Thirty-two Sprague-Dawley rats were randomly assigned to four different treatment groups. One group served as the non-diabetic control, while diabetes was induced in other groups. Titanium plasma-sprayed (TPS) implants were placed in the femora of each animal 2 weeks following diabetic induction. One group of diabetic rats was given aminoguanidine via intraperitoneal injection, and another given doxycycline via oral gavage for 28 days beginning on the day of implantation. The third group of diabetic rats received no medication (controls). All animals were sacrificed following 28 days of healing. RESULTS: The results were measured by marrow bone-to implant contact (MBIC) between the groups. Values for MBIC were greater for the non-diabetic control group than the diabetic control group (P < 0.001). Aminoguanidine-treated diabetic animals had a significantly greater MBIC than the diabetic control group (P < 0.01). Diabetic animals receiving doxycycline did not differ significantly from the diabetic control group (P > 0.05). CONCLUSIONS: The results of this study using a rat model con- firm previous reports that diabetes inhibits osseointegration, as defined by MBIC. In addition, this study demonstrates that the detrimental effects of diabetes on osseointegration can be modified using aminoguanidine systemically. However, systemic administration of doxycycline only slightly enhances osseointegration.

Maintenance of osseointegration utilizing insulin therapy in a diabetic rat model.

BACKGROUND: Normal wound healing processes have been shown to be altered in diabetes, and the effect of the diabetes on bone-to-implant contact (BIC) once osseointegration has been established is still unknown. The purpose of this study was to histologically evaluate the bone-to-implant contact in uncontrolled and insulin-controlled rats in which diabetes was induced following the establishment of osseointegration. METHODS: Thirty-two rats were assigned to eight different treatment groups of four each. Titanium plasma-sprayed (TPS) implants were placed in the femora of each animal, and allowed to osseointegrate for 28 days before diabetic induction. Daily insulin injections were given to four groups of rats and the other four groups received no insulin (uncontrolled). The rats were sacrificed at 1, 2, 3, and 4 months following diabetic induction. RESULTS: The results indicated that at 1, 2, 3, and 4 months, there was more BIC in the insulin-controlled groups compared to the uncontrolled groups. The differences were significantly greater at 2, 3, and 4 months (P < or =0.001). CONCLUSIONS: This study demonstrated that osseointegrated dental implants in insulin-controlled diabetic rats maintained bone-to-implant contacts over a 4-month period. However, boneto- implant contact appears to decrease with time in uncontrolled diabetic rats.

Accuracy of administrative billing codes to detect urinary tract infection hospitalizations.

BACKGROUND: Hospital billing data are frequently used for quality measures and research, but the accuracy of the use of discharge codes to identify urinary tract infections (UTIs) is unknown. OBJECTIVE: To determine the accuracy of International Classification of Diseases, 9th revision (ICD-9) discharge codes to identify children hospitalized with UTIs. METHODS: This multicenter study conducted in 5 children's hospitals included children aged 3 days to 18 years who had been admitted to the hospital, undergone a urinalysis or urine culture, and discharged from the hospital. Data were obtained from the pediatric health information system database and medical record review. With the use of 2 gold-standard methods, the positive predictive value (PPV) was calculated for individual and combined UTI codes and for common UTI identification strategies. PPV was measured for all groupings for which the UTI code was the principal discharge diagnosis. RESULTS: There were 833 patients in the study. The PPV was 50.3% with the use of the gold standard of laboratory-confirmed UTIs but increased to 85% with provider confirmation. Restriction of the study cohort to patients with a principle diagnosis of UTI improved the PPV for laboratory-confirmed UTI (61.2%) and provider-confirmed UTI (93.2%), as well as the ability to benchmark performance. Other common identification strategies did not markedly affect the PPV. CONCLUSIONS: ICD-9 codes can be used to identify patients with UTIs but are most accurate when UTI is the principal discharge diagnosis. The identification strategies reported in this study can be used to improve the accuracy and applicability of benchmarking measures.