Evaluation of Two Multiplex PCR-High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies.

Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)-high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene. However, higher intraspecies variation within C. coli and C. jejuni isolates was detected in mPCR1-HRM when compared with mPCR2-HRM. Both assays were rapid and required minimum interpretation skills for discrimination between and within Campylobacter species when using HRM curve analysis software.

Differentiation of Campylobacter jejuni and Campylobacter coli Using Multiplex-PCR and High Resolution Melt Curve Analysis.

Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates.

Sex preference as a determinant of contraceptive use in matrilineal societies: a study on the Garo of Bangladesh.

OBJECTIVES: This paper explores the presence of sex preference and its influence on contraceptive use among the Garo, a matrilineal community, of Bangladesh. METHODS: In this study, 223 currently married Garo women were interviewed, selected purposively from two districts of Bangladesh, where most of the Garo people live. A binary logistic regression model was fitted to explore the determinants of current use of contraceptives among the Garo community. RESULTS: About 80% of the Garo women were currently using contraceptive methods, which was much higher than the contraceptive prevalence at the national level (55.8%). However, the use of modern male methods was very low in that community. Regression analysis revealed that the strong preference for girls was a significant determinant of contraceptive use. CONCLUSIONS: Even though the Garo apply contraception more than the Bangladeshi community in general, their strong preference for girls may restrain its success. Family planning information can be designed so as not to challenge the matrilineal structure of the society and both spouses counselled together to promote also modern male methods.

Determinants of exposure to mass media family planning messages among indigenous people in Bangladesh: a study on the Garo.

This paper evaluates exposure to mass media family planning (FP) messages among the Garo, an indigenous community in Bangladesh. A sample of 223 currently married Garo women were selected purposively from two districts where most of the Garo population live. The analysis demonstrated that television was the most significant form of mass media to disseminate FP messages among the recipients - more so than radio and newspapers. About 80.6% of the respondents had heard of FP messages through television, while for the radio and newspapers the percentages were 55.3% and 22.7% respectively. The contraceptive prevalence rate is much higher (79.5%) in the study area than the national level (55.8%). A linear logistic regression model was employed to identify the confluence of different demographic and socioeconomic characteristics on mass media FP messages. Regarding exposure to FP messages, four independent variables out of six had significant effects on the exposure to FP messages through any one of the types of media, i.e. radio, television and newspapers. These independent variables were age, level of education, occupation and number of children.