A comparative study was performed to evaluate best practice culture media and enrichment broths for recovering Salmonella species from human stool samples. A total of 1297 human stools were collected and processed in ...A comparative study was performed to evaluate best practice culture media and enrichment broths for recovering Salmonella species from human stool samples. A total of 1297 human stools were collected and processed in this study. Evaluation of agar media was carried out by direct plating (DP), 1096 stool samples were inoculated on Modified Semisolid Rappaport-Vassiliadis (MSRV), Xylose-Lysine-Deoxycolate (XLD), MacConkey (MAC), and Hektoen Enteric (HE) agars. Evaluation of enrichment broths were carried out by enrichment all 1297 stool samples in Selenite broth (SB), Rappaport-Vassiliadis (RV) broth, and Buffered Peptone Water (BPW), followed by plating on MSRV, MAC, and HE agars. A total of 102 Salmonella-positive stools by DP, 85.3% (87/102) were recovered utilizing MSRV while recovery from XLD, MAC, and HE agars were 34.3% (35/102), 34.3% (35/102), and 29.4% (30/102) respectively. A total 299/1297 stools samples were Salmonella-positive on at least one plating medium after enrichment procedure were 77.3% (177/299) for SB, 86.0% (197/299) and 78.6% (180/299) for RV and BPW respectively. All Salmonella isolated in this study was nontyphi Salmonella. Presently, the data suggest that the use of MSRV over MAC, HE, and XLD agars for isolation nontyphi Salmonella species from human stools is more efficacious. Additionally, use of MSRV in combination with MAC and HE agars following enrichment in RV broth enhances recovery of nontyphi Salmonella species. However, RV broth is inhibitory to typhi Salmonella, thus use of MSRV medium in combination with MAC, HE or XLD agars in direct plating following enrichment in non-selective BPW is an alternate method for recovery of both typhi and nontyphi Salmonella species contaminated in human stool samples.展开更多
Campylobacter continues to be a major cause of bacteriamediated diarrheal diseases, both for Thai citizens and travelers to Thailand. For field epidemiological studies, appropriate methods for storage, intralaboratory...Campylobacter continues to be a major cause of bacteriamediated diarrheal diseases, both for Thai citizens and travelers to Thailand. For field epidemiological studies, appropriate methods for storage, intralaboratory transport of patients specimens and use of enrichment culture to isolate this organism is critical. Study A, represents patient stool specimens collected in Bangkok and processed for Campylobacter culture within three hours after collection. Study B, represents stool specimens collected from patients in northeast and Southern regions of Thailand in modified CaryBlair transport medium. These specimens were transported and processed for Campylobacter in Bangkok at varying intervals ranging from 1 to 7 days. Of 900 diarrheal samples examined in study A, a total of 158 were Campylobacter positive through culture. Of these, 145 and 141 isolates were cultured by direct plating and enrichment plating respectively (P = 0.5839). From 1,168 diarrheal stool samples examined in study B, 184 were positive for Campylobacter. Direct and enrichment plating resulted in 139 and 168 culture isolates;respectively (P = 0.0003). Samples from study B delayed in processing for 1 to 3 days, resulted in 46 and 50 isolated by direct and enrichment plating;respectively (P = 0.4545). However, among samples delayed in processing for 4 to 7 days, a total of 128 Campylobacter isolates were cultured, having cultured 93 and 118 isolates through direct and enrichment plating;respectively (P = 0.0003). At present these studies demonstrate that enrichment culture has no benefit when stool specimen collection and immediate processing occur and when there is a processing delay period of 1 - 3 days. However, enrichment culture was beneficial in instances where transport and processing was delayed 4 - 7 days.展开更多
目的探讨不同贮存温度下临床粪便标本中艰难梭菌毒素的免疫学稳定性。方法应用酶免疫分析(enzyme immunoassay,EIA)试剂盒检测定量聚合酶链式反应确认为艰难梭菌感染(Clostridium di f ficile infection,CDI)的粪便标本中艰难梭菌毒素...目的探讨不同贮存温度下临床粪便标本中艰难梭菌毒素的免疫学稳定性。方法应用酶免疫分析(enzyme immunoassay,EIA)试剂盒检测定量聚合酶链式反应确认为艰难梭菌感染(Clostridium di f ficile infection,CDI)的粪便标本中艰难梭菌毒素含量。将EIA阳性标本置于4、-20、-80℃有氧条件下储存,在储存的第1、14、30、60、90、120天通过EIA法测定毒素含量。结果共收集43例患者粪便标本,定量聚合酶链式反应检测出CDI阳性标本7例(16.28%),其中4例EIA检测为艰难梭菌毒素阳性,毒素含量差异较大;未使用储存缓冲液的粪便标本中艰难梭菌毒素稳定性较好,相同温度下使用储存缓冲液的粪便标本中艰难梭菌毒素稳定性较差、降解较快,-20℃时稳定性最差。结论不同储存温度对未使用储存缓冲液的临床粪便标本艰难梭菌毒素的免疫稳定性没有明显影响。但稀释保存不利于毒素稳定性,尤其是-20℃,临床中应避免使用该储存温度。展开更多
文摘A comparative study was performed to evaluate best practice culture media and enrichment broths for recovering Salmonella species from human stool samples. A total of 1297 human stools were collected and processed in this study. Evaluation of agar media was carried out by direct plating (DP), 1096 stool samples were inoculated on Modified Semisolid Rappaport-Vassiliadis (MSRV), Xylose-Lysine-Deoxycolate (XLD), MacConkey (MAC), and Hektoen Enteric (HE) agars. Evaluation of enrichment broths were carried out by enrichment all 1297 stool samples in Selenite broth (SB), Rappaport-Vassiliadis (RV) broth, and Buffered Peptone Water (BPW), followed by plating on MSRV, MAC, and HE agars. A total of 102 Salmonella-positive stools by DP, 85.3% (87/102) were recovered utilizing MSRV while recovery from XLD, MAC, and HE agars were 34.3% (35/102), 34.3% (35/102), and 29.4% (30/102) respectively. A total 299/1297 stools samples were Salmonella-positive on at least one plating medium after enrichment procedure were 77.3% (177/299) for SB, 86.0% (197/299) and 78.6% (180/299) for RV and BPW respectively. All Salmonella isolated in this study was nontyphi Salmonella. Presently, the data suggest that the use of MSRV over MAC, HE, and XLD agars for isolation nontyphi Salmonella species from human stools is more efficacious. Additionally, use of MSRV in combination with MAC and HE agars following enrichment in RV broth enhances recovery of nontyphi Salmonella species. However, RV broth is inhibitory to typhi Salmonella, thus use of MSRV medium in combination with MAC, HE or XLD agars in direct plating following enrichment in non-selective BPW is an alternate method for recovery of both typhi and nontyphi Salmonella species contaminated in human stool samples.
文摘Campylobacter continues to be a major cause of bacteriamediated diarrheal diseases, both for Thai citizens and travelers to Thailand. For field epidemiological studies, appropriate methods for storage, intralaboratory transport of patients specimens and use of enrichment culture to isolate this organism is critical. Study A, represents patient stool specimens collected in Bangkok and processed for Campylobacter culture within three hours after collection. Study B, represents stool specimens collected from patients in northeast and Southern regions of Thailand in modified CaryBlair transport medium. These specimens were transported and processed for Campylobacter in Bangkok at varying intervals ranging from 1 to 7 days. Of 900 diarrheal samples examined in study A, a total of 158 were Campylobacter positive through culture. Of these, 145 and 141 isolates were cultured by direct plating and enrichment plating respectively (P = 0.5839). From 1,168 diarrheal stool samples examined in study B, 184 were positive for Campylobacter. Direct and enrichment plating resulted in 139 and 168 culture isolates;respectively (P = 0.0003). Samples from study B delayed in processing for 1 to 3 days, resulted in 46 and 50 isolated by direct and enrichment plating;respectively (P = 0.4545). However, among samples delayed in processing for 4 to 7 days, a total of 128 Campylobacter isolates were cultured, having cultured 93 and 118 isolates through direct and enrichment plating;respectively (P = 0.0003). At present these studies demonstrate that enrichment culture has no benefit when stool specimen collection and immediate processing occur and when there is a processing delay period of 1 - 3 days. However, enrichment culture was beneficial in instances where transport and processing was delayed 4 - 7 days.
文摘目的探讨不同贮存温度下临床粪便标本中艰难梭菌毒素的免疫学稳定性。方法应用酶免疫分析(enzyme immunoassay,EIA)试剂盒检测定量聚合酶链式反应确认为艰难梭菌感染(Clostridium di f ficile infection,CDI)的粪便标本中艰难梭菌毒素含量。将EIA阳性标本置于4、-20、-80℃有氧条件下储存,在储存的第1、14、30、60、90、120天通过EIA法测定毒素含量。结果共收集43例患者粪便标本,定量聚合酶链式反应检测出CDI阳性标本7例(16.28%),其中4例EIA检测为艰难梭菌毒素阳性,毒素含量差异较大;未使用储存缓冲液的粪便标本中艰难梭菌毒素稳定性较好,相同温度下使用储存缓冲液的粪便标本中艰难梭菌毒素稳定性较差、降解较快,-20℃时稳定性最差。结论不同储存温度对未使用储存缓冲液的临床粪便标本艰难梭菌毒素的免疫稳定性没有明显影响。但稀释保存不利于毒素稳定性,尤其是-20℃,临床中应避免使用该储存温度。