Drug sensitivity testing to establish resistance to TB drugs takes many months to arrive at. Public health physicians have difficulties with such an approach due to long wait periods and cannot use it to establish com...Drug sensitivity testing to establish resistance to TB drugs takes many months to arrive at. Public health physicians have difficulties with such an approach due to long wait periods and cannot use it to establish community wide prevalence as a way to understand where resistance may be emerging faster and to limit its spread. The objective of this study was to use the dot-blot hybridization technique in the detection of resistance to rifamycin (RIF) and streptomycin (SM) in South- Western Cameroon and to compare the technique with the routine culture and drug susceptibility testing for detecting resistance in a resource poor country, Cameroon. A hospital-based study was conducted at the Regional hospitals of Buea and Limbe and Tiko Central Clinic. Tuberculosis (TB) patients aged 15 to 50 (mean age: 30.50 ± 8.33 standard deviation) were recruited for the study between December 2006 and April 2007. Cultures from 59 patients were tested for rifampicin and streptomycin sensitivity by the modified proportion method and mutational analysis for rpoB codon 516 and rrs codon 513 was performed by the dot-blot hybridization technique. Of the 59 sputum samples collected (36 were males and 23 were females) came from Buea 19 (32.2%), Limbe 20 (33.9%) and Tiko 20 (33.9%) towns respectively. Amplification for the gene showed that there was (59) 100% amplification with primers used for rpoB genes and 43 (72.9%) amplification with primers used for the rrs gene. Mutational analysis demonstrated that resistance to RIF was common in females (52.1%) than males (41.7%) while 6% of the samples were indeterminate. 12 (20.3%) samples showed phenotypic and genotypic resistance to RIF compared to 34 samples (58.1%) for SM. Phenotypic resistance and genotypic susceptibility were found in 5 (8.5%) RIF and 3 (4.7%) SM compared to phenotypic susceptibility and genotypic resistance that were found in 2 (3.5%) RIF and 3(4.7%) SM. Double mutation on rpoB and rrs genes occurred in 8 (13.6%) DNA samples. Resistance to RIF and SM due to mutations on the rpoB and rrs genes respectively in the SW region was found to be high and comparable to the drug susceptibility testing by 92%, (95% CI: 75.7 - 99.1). The Dot-blot technique will be useful in rapidly assessing the effectiveness of national TB control programs in limiting the spread of resistance strains in Cameroon.展开更多
Meat constitutes the main source of protein and occupies an important place in our diet. Indeed, the production of poultry and beef has increased. However, the hygienic quality of meat is not always guaranteed. Microo...Meat constitutes the main source of protein and occupies an important place in our diet. Indeed, the production of poultry and beef has increased. However, the hygienic quality of meat is not always guaranteed. Microorganisms such as Escherichia coli can be found in meat and can cause various infections including diarrhea, dysentery, food poisoning, gastroenteritis or typhoid fever. Thus, the present study was designed to characterize Escherichia coli (E. coli) from beef and chicken consumed in restaurants in Yaoundé Cameroon. A total of 105 meat samples (60 beef and 45 chickens) were subjected to microbial culture for E. coli isolation and further confirmed by Polymerase Chain Reaction (PCR) using primers EC-F and EC-R that are specific to E. coli 16S rRNA gene. The supplier source, storage, and transport conditions were taken into consideration during sample analysis and data processing. This study revealed that 77/105 samples (73.33%) were positive for E. coli following microbial culture and 35 (33.33%) were positive for E. coli following molecular examination. A statistically significant difference was observed when PCR and microbial culture were used to assess for E. coli in beef and a non-statistically significant difference was observed in the case of chicken meat. Also, a statistically significant difference was noticed with the different transport conditions, but this wasn’t the case with the supplier source as well as the storage conditions where a non-statistically significant difference was seen. This study revealed that PCR-based methods are fast and reliable in the identification and characterization of Escherichia coli in meats (beef and chicken) as well as in assessing the prevalence of pathogenic E. coli, in Cameroon.展开更多
文摘Drug sensitivity testing to establish resistance to TB drugs takes many months to arrive at. Public health physicians have difficulties with such an approach due to long wait periods and cannot use it to establish community wide prevalence as a way to understand where resistance may be emerging faster and to limit its spread. The objective of this study was to use the dot-blot hybridization technique in the detection of resistance to rifamycin (RIF) and streptomycin (SM) in South- Western Cameroon and to compare the technique with the routine culture and drug susceptibility testing for detecting resistance in a resource poor country, Cameroon. A hospital-based study was conducted at the Regional hospitals of Buea and Limbe and Tiko Central Clinic. Tuberculosis (TB) patients aged 15 to 50 (mean age: 30.50 ± 8.33 standard deviation) were recruited for the study between December 2006 and April 2007. Cultures from 59 patients were tested for rifampicin and streptomycin sensitivity by the modified proportion method and mutational analysis for rpoB codon 516 and rrs codon 513 was performed by the dot-blot hybridization technique. Of the 59 sputum samples collected (36 were males and 23 were females) came from Buea 19 (32.2%), Limbe 20 (33.9%) and Tiko 20 (33.9%) towns respectively. Amplification for the gene showed that there was (59) 100% amplification with primers used for rpoB genes and 43 (72.9%) amplification with primers used for the rrs gene. Mutational analysis demonstrated that resistance to RIF was common in females (52.1%) than males (41.7%) while 6% of the samples were indeterminate. 12 (20.3%) samples showed phenotypic and genotypic resistance to RIF compared to 34 samples (58.1%) for SM. Phenotypic resistance and genotypic susceptibility were found in 5 (8.5%) RIF and 3 (4.7%) SM compared to phenotypic susceptibility and genotypic resistance that were found in 2 (3.5%) RIF and 3(4.7%) SM. Double mutation on rpoB and rrs genes occurred in 8 (13.6%) DNA samples. Resistance to RIF and SM due to mutations on the rpoB and rrs genes respectively in the SW region was found to be high and comparable to the drug susceptibility testing by 92%, (95% CI: 75.7 - 99.1). The Dot-blot technique will be useful in rapidly assessing the effectiveness of national TB control programs in limiting the spread of resistance strains in Cameroon.
文摘Meat constitutes the main source of protein and occupies an important place in our diet. Indeed, the production of poultry and beef has increased. However, the hygienic quality of meat is not always guaranteed. Microorganisms such as Escherichia coli can be found in meat and can cause various infections including diarrhea, dysentery, food poisoning, gastroenteritis or typhoid fever. Thus, the present study was designed to characterize Escherichia coli (E. coli) from beef and chicken consumed in restaurants in Yaoundé Cameroon. A total of 105 meat samples (60 beef and 45 chickens) were subjected to microbial culture for E. coli isolation and further confirmed by Polymerase Chain Reaction (PCR) using primers EC-F and EC-R that are specific to E. coli 16S rRNA gene. The supplier source, storage, and transport conditions were taken into consideration during sample analysis and data processing. This study revealed that 77/105 samples (73.33%) were positive for E. coli following microbial culture and 35 (33.33%) were positive for E. coli following molecular examination. A statistically significant difference was observed when PCR and microbial culture were used to assess for E. coli in beef and a non-statistically significant difference was observed in the case of chicken meat. Also, a statistically significant difference was noticed with the different transport conditions, but this wasn’t the case with the supplier source as well as the storage conditions where a non-statistically significant difference was seen. This study revealed that PCR-based methods are fast and reliable in the identification and characterization of Escherichia coli in meats (beef and chicken) as well as in assessing the prevalence of pathogenic E. coli, in Cameroon.
