Inactivation of Escherichia Coli O157:H7 and Salmonella Enterica on Blueberries in Water Using Ultraviolet Light

Ultraviolet light (UV) has antimicrobial effects, but the shadowing effect has limited its application. In this study, a novel setup using UV processing in agitated water was developed to inactivate Escherichia coli O157:H7 and Salmonella on blueberries. Blueberries were dip‐ or spot‐inoculated with E. coli or Salmonella. Blueberries inoculated with E. coli were treated for 2 to 10 min with UV directly (dry UV) or immersed in agitated water during UV treatment (wet UV). E. coli was most easily killed on spot‐inoculated blueberries with a 5.2‐log reduction after 10‐min wet UV treatment. Dip‐inoculated blueberries were the most difficult to be decontaminated with only 1.6‐log reduction after 10‐min wet UV treatment. Wet UV treatment generally showed higher efficacies than dry UV treatment, achieving an average of 1.4 log more reduction for spot‐inoculated blueberries. For dip‐inoculated blueberries, chlorine washing and UV treatments were less effective, achieving <2 log reductions of E. coli. Thus, the efficacy of combinations of wet UV with sodium dodecyl sulfate (SDS), levulinic acid, or chlorine was evaluated. Inoculated blueberries were UV‐treated while being immersed in agitated water containing 100 ppm SDS, 0.5% levulinic acid or 10 ppm chlorine. The 3 chemicals did not significantly enhance the wet UV treatment. Findings of this study suggest that UV treatment could be used as an alternative to chlorine washing for blueberries and potentially for other fresh produce.     

特异性抗肠出血性大肠杆菌O157:H7内毒素鸡卵黄抗体的制备

目的:比较研究LPS和O-SP的免疫原性,制备出特异性抗内毒素(LPS)鸡卵黄免疫球蛋白(eggyolkimmunoglobulin,IgY),用于对E.coliO157:H7的免疫预防及检测。方法:分别用灭活E.coliO157:H7菌体、不同浓度内毒素(LPS)及O-特异性多糖链(O-SP)与不完全弗氏佐剂充分乳化后作抗原免疫临产蛋母鸡,以水稀释法从卵黄中提取抗体,阴离子交换色谱法实现对抗体的纯化,间接ELISA及双向免疫扩散检测抗体产生效价与活性。结果与结论:所制内毒素(LPS)和O-SP均有较好的免疫原性,刺激鸡体后可产生高效价抗体,灭活菌体、600μg/mlLPS、1200μg/mlLPS、2000μg/mlLPS、O-SP抗体产生效价最高可分别达1:32000、1:28000、1:32000、1:12000、1:40000。结合离子交换色谱,用0.185mol/L的离子强度可实现一步洗脱纯化抗体,制备出高纯度、高活性、特异性强的鸡卵黄免疫球蛋白IgY,为大肠杆菌O157:H7的感染防治提供了可靠的保证,在此实验基础之上可进一步探讨应用特异性IgY对大肠杆菌O157:H7的检测。     

Efficacy of Ozone Against Escherichia coli O157:H7 on Apples

Apples were inoculated with Escherichia coli O157:H7 and treated with ozone. Sanitization treatments were more effective when ozone was bubbled during apple washing than by dipping apples in pre‐ozonated water. The corresponding decreases in counts of E. coli O157:H7 during 3‐min treatments were 3.7 and 2.6 log₁₀ CFU on apple surface, respectively, compared to < 1 log₁₀ CFU decrease in the stem‐calyx region in both delivery methods. Optimum conditions for decontamination of whole apples with ozone included a pretreatment with a wetting agent, followed by bubbling ozone for 3 min in the wash water, which decreased the count of E. coli O157:H7 by 3.3 log₁₀CFU/g.     

Response Surface Modeling for the Inactivation of Escherichia coli O157:H7 on Green Peppers (Capsicum annuum) by Ozone Gas Treatment

ABSTRACT: :
The effects of ozone gas concentration (2 to 8 mg/l), relative humidity (RH) (60 to 90%), and treatment time (10 to 40 min) on inactivation of E. coli O157:H7 on green peppers were studied using response surface methodology. A 3-factor Box-Behnken experimental plan was designed and microbial log reduction was measured as a response. The statistical analysis of developed predictive model suggested that ozone gas concentration, RH, and treatment time all significantly (P < 0.01) increased the rate of log reduction of E. coli O157:H7. Among the 3 factors, the effect of ozone gas concentration on bacterial inactivation was the greatest, while the effect of RH was the least. The interaction between ozone gas concentration and RH exhibited a significant and synergistic effect (P < 0.05).     

Inactivation of E. coli O157:H7 Using a Pulsed Nonthermal Plasma System

ABSTRACT: The use of elevated temperatures to pasteurize food products leads to a loss of food quality, which stimulates research into novel nonthermal processes for food preservation. Using nonthermal plasma discharges, applied directly into the food product, proved effective in reducing the number of Escherichia coli O157:H7 cells in apple juice, by up to 7 log units. The pulsed nonthermal plasma system was developed and used to study the effects of different process parameters on the inactivation. The process parameters considered were discharge voltage, pulse number, and pulse frequency. The apple juice used had a pH of 4 and a conductivity of 2.29 mS/cm.     

Survival and Growth of Vibrio cholerae O1, Salmonella typhi, and Escherichia coli O157:H7 in Alfalfa Sprouts

The survival and growth of Vibrio cholerae O1, Salmonella typhi , and Escherichia coli O157:H7 during germination and sprouting of disinfected alfalfa seeds and alfalfa sprouts was determined. All pathogens showed ability to grow during germination and sprouting, reaching counts of approximately 6.0 log₁₀ CFU/g after 24 h. No growth was observed for any pathogen when the sprouts were inoculated after 24 h of seed germination. At this time, the background microflora was abundant. Numbers of pathogens inoculated on alfalfa sprouts decreased less than 1 log10 CFU/g over 15 d of refrigeration. Alfalfa sprouts can be an important factor contributing to the endemicity for typhoid fever and cholera in México.     

Comparison of Decontamination Efficacy of Antimicrobial Treatments for Beef Trimmings against Escherichia coli O157:H7 and 6 Non-O157 Shiga Toxin-Producing E. coli Serogroups

Abstract: The decontamination efficacy of 6 chemical treatments for beef trimmings were evaluated against Escherichia coli O157:H7 and 6 non‐O157 Shiga toxin‐producing E. coli (nSTEC) serogroups. Rifampicin‐resistant 4‐strain mixtures of E. coli O157:H7 and nSTEC serogroups O26, O45, O103, O111, O121, and O145 were separately inoculated (3 to 4 log CFU/cm²) onto trimmings (10 × 5 × 1 cm; approximately 100 g) fabricated from beef chuck rolls, and were immersed for 30 s in solutions of acidified sodium chlorite (0.1%, pH 2.5), peroxyacetic acid (0.02%, pH 3.8), sodium metasilicate (4%, pH 12.5), Bromitize^® Plus (0.0225% active bromine, pH 6.6), or AFTEC 3000 (pH 1.2), or for 5 s in SYNTRx 3300 (pH 1.0). Each antimicrobial was tested independently together with an untreated control. Results showed that all tested decontamination treatments were similarly effective against the 6 nSTEC serogroups as they were against E. coli O157:H7. Irrespective of pathogen inoculum, treatment of beef trimmings with acidified sodium chlorite, peroxyacetic acid, or sodium metasilicate effectively (P < 0.05) reduced initial pathogen counts (3.4 to 3.9 log CFU/cm²) by 0.7 to 1.0, 0.6 to 1.0, and 1.3 to 1.5 log CFU/cm², respectively. Reductions of pathogen counts (3.1 to 3.2 log CFU/cm²) by Bromitize Plus, AFTEC 3000, and SYNTRx 3300 were 0.1 to 0.4 log CFU/cm², depending on treatment. Findings of this study should be useful to regulatory authorities and the meat industry as they consider nSTEC contamination in beef trimmings. Practical Applications: Findings of this study should be useful to: (i) meat processors as they design and conduct studies to validate the efficacy of antimicrobial treatments to control pathogen contamination on fresh beef products; and (ii) regulatory agencies as they consider approaches for better control of the studied pathogens.     

Inactivation of Escherichia coli O157:H7 and Salmonella enteritidis in Liquid Egg White Using Pulsed Electric Field

ABSTRACT: The effects of temperature and pulsed electric field (PEF) intensity on inactivation of pathogens such as Escherichia coli O157:H7 and Salmonella enteritidis in egg white was investigated. Liquid egg white inoculated with 10⁸ colony-forming units (CFU)/mL of each pathogen was treated with up to 60 pulses (each of 2 JAS width) at electric field intensities of 20 and 30 kV/cm. The processing temperatures were 10°C, 20°C, and 30°C. After treatment, uninjured and total viable cells were enumerated in selective and nonselective agars, respectively. Maximum inactivations of 3.7 and 2.9 log units were obtained for S. enteritidis and E. coli O157:H7, respectively, while injured cells accounted for 0.5 and 0.9 logs for E. coli O157:H7 and S. enteritidis , respectively. For both bacteria, increasing treatment temperature tended to increase the inactivation rate. There was synergy between electric field intensity and processing temperature. The inactivation rate constant k _T values for E. coli O157:H7 on both selective and nonselective agars were 8.2 × 10^-3 and 6.6 × 10^-3/μS, whereas the values for S. enteritidis were 16.2 × 10^-3 and 12.6 × 10^-3/μS, respectively. The results suggest that E. coli O157:H7 was more resistant to heat-PEF treatment compared with S. enteritidis.     

Efficacy of Integrated Treatment of UV light and Low‐Dose Gamma Irradiation on Inactivation of Escherichia coli O157:H7 and Salmonella enterica on Grape Tomatoes

The study evaluated the efficacy of integrated ultraviolet‐C light (UVC) and low‐dose gamma irradiation treatments to inactivate mixed strains of Escherichia coli O157:H7 and Salmonella enterica on inoculated whole grape tomatoes. A mixed bacterial cocktail composed of a 3 strain mixture of E. coli O157:H7 (C9490, E02128, and F00475) and a 3 serotype mixture of S. enterica (S. Montevideo G4639, S. Newport H1275, and S. Stanley H0558) was used based on their association with produce‐related outbreaks. Spot inoculation (50 to 100 μmL) on tomato surfaces was performed to achieve a population of appropriately 10^7–8 CFU/tomato. Inoculated tomatoes were subjected to UVC (253.7 nm) dose of 0.6 kJ/m² followed by 4 different low doses of gamma irradiations (0.1 kGy, 0.25 kGy, 0.5 kGy, 0.75 kGy). The fate of background microflora (mesophilic aerobic) including mold and yeast counts were also determined during storage at 5 °C over 21 d. Integrated treatment significantly (P < 0.05) reduced the population of target pathogens. Results indicate about 3.4 ± 0.3 and 3.0 ± 0.1 log CFU reduction of E. coli O157:H7 and S. enterica, respectively, per tomato with UVC (0.6 kJ/m²) and 0.25 kGy irradiation. More than a 4 log and higher reduction (>5 log) per fruit was accomplished by combined UVC treatment with 0.5 kGy and 0.75 kGy irradiation, respectively, for all tested pathogens. Furthermore, the combined treatment significantly (P < 0.05) reduced the native microflora compared to the control during storage. The data suggest efficacious treatment strategy for produce indicating 5 or higher log reduction which is consistent with the recommendations of the Natl. Advisory Committee on Microbiological Criteria for Foods.     

A note on Escherichia coli O157:H7 serotype in Turkish meat products

255 minced beef, 101 soudjouk and 50 uncooked hamburger samples were analyzed for the presence of Escherichia coli O157:H7 serotype. m-EC and LST broths were used as selective enrichment media and SMAC agar was used as a selective isolation medium. A total of 3 E. coli O157 were isolated by conventional culture techniques; one from each of minced beef, uncooked hamburger and soudjouk but none were identified as the H7 serotype. For determination of selective media-growing cohabitant bacteria, 2645 isolates were obtained from SMAC agar. Results showed that E. coli type 1, Hafnia alvei and Citrobacter freundii were dominant competitive flora. As selective enrichment broth-growing cohabitant microflora existed at higher levels, it was too difficult to isolate E. coli O157 from these mixed flora. Therefore, conventional methods are not suitable for these types of products, because of isolation difficulties and failure to confirm.     

Survival of Escherichia coli O157:H7 in cucumber fermentation brines

Abstract: Bacterial pathogens have been reported on fresh cucumbers and other vegetables used for commercial fermentation. The Food and Drug Administration currently has a 5‐log reduction standard for E. coli O157:H7 and other vegetative pathogens in acidified pickle products. For fermented vegetables, which are acid foods, there is little data documenting the conditions needed to kill acid resistant pathogens. To address this knowledge gap, we obtained 10 different cucumber fermentation brines at different stages of fermentation from 5 domestic commercial plants. Cucumber brines were used to represent vegetable fermentations because cabbage and other vegetables may have inhibitory compounds that may affect survival. The 5‐log reduction times for E. coli O157:H7 strains in the commercial brines were found to be positively correlated with brine pH, and ranged from 3 to 24 d for pH values of 3.2 to 4.6, respectively. In a laboratory cucumber juice medium that had been previously fermented with Lactobacillus plantarum or Leuconostoc mesenteroides (pH 3.9), a 5‐log reduction was achieved within 1 to 16 d depending on pH, acid concentration, and temperature. During competitive growth at 30 °C in the presence of L. plantarum or L. mesenteroides in cucumber juice, E. coli O157:H7 cell numbers were reduced to below the level of detection within 2 to 3 d. These data may be used to aid manufacturers of fermented vegetable products determine safe production practices based on fermentation pH and temperature. Practical Application: Disease causing strains of the bacterium E. coli may be present on fresh vegetables. Our investigation determined the time needed to kill E. coli in cucumber fermentation brines and how E. coli strains are killed in competition with naturally present lactic acid bacteria. Our results showed how brine pH and other brine conditions affected the killing of E. coli strains. These data can be used by producers of fermented vegetable products to help assure consumer safety.     

Statistical Modeling of D- and z-value of E. coli O157:H7 and pH in Apple Cider Containing Preservatives

ABSTRACT: The objective of this study was to model the heat resistance of E. coli O157:H7 in apple cider as a function of added sorbate, benzoate, and malic acid. Inactivation experiments were done at 47, 50, and 53 °C, and D and z values were calculated. Models for D₄₇, D₅₀, and D₅₃ showed that all 3 additives significantly decreased heat resistance. Furthermore, z-values increased with addition of preservatives and low pH. Model predictions showed that at 50 °C, 5-log reduction is attainable in a few minutes when the cider contains additives, while preliminary test results showed that at 20 °C, a 5-log reduction could be obtained in about 90 min in cider with 0.1% sorbate, 0.1% benzoate, and 1% malic acid.     

Thermal Inactivation of Escherichia coli O157:H7 in Strawberry Puree and Its Effect on Anthocyanins and Color

Raw whole strawberries, if contaminated with pathogens, such as Escherichia coli O157:H7, must be pasteurized prior to consumption. Therefore, the objective of this research was to investigate the thermal inactivation kinetics of E. coli O157:H7 in strawberry puree (SP), and evaluate the changes in anthocyanins and color, and the survival of yeasts and molds (YM) after thermal processing. Inoculated with a 5‐strain cocktail, fresh SP, with or without added sugar (20 and 40 °Brix), was heated at 50, 52, 54, 57.5, 60, and 62.5 °C to determine the thermal resistance of E. coli O157:H7. In raw SP, the average D‐values of E. coli O157:H7 were 909.1, 454.6, 212.8, 46.1, and 20.2 s at 50, 52, 54, 57.5, and 60 °C, respectively, with a z‐value of 5.9 °C. While linearly decreasing with temperature, the log D‐values of E. coli O157:H7 increased slightly with sugar concentration. The log degradation rates of anthocyanins increased linearly with temperature, but decreased slightly with sugar concentrations. These results suggest that sugar may provide some protection to both E. coli O157: H7 and anthocyanins in SP. The browning index was not affected by heating at 50 and 52 ºC at low sugar concentrations, but increased by an average of 1.28%, 2.21%, and 10.1% per min when SP was exposed to heating at 54, 57.5, and 60 °C, respectively. YM was also inactivated by heating. This study demonstrated that properly designed thermal processes can effectively inactivate E. coli O157:H7 and YM in contaminated SP, while minimizing the changes in anthocyanins and color.     

Injury,Inhibition and Inactivation ofEscherichia coli O157:H7 by Potassium Sorbate and Sodium Nitrite as Affected by pH and Temperature

The effect of potassium sorbate (0–2 g litre⁻¹) and sodium nitrite (0–1 g litre⁻¹) on the growth of four strains of Escherichia coli O157: H7 in tryptic soya broth at various pH levels (pH 4·0–7·0 for sorbate, pH 5·0–8·0 for nitrite) were determined at 37°C and 4°C. Among the pH levels tested, sorbate and nitrite exhibited the highest antimicrobial activity at pH 4·0 and 5·0, respectively. At pH 5·0 and 37°C, the presence of 500 mg litre⁻¹ sorbate or 200 mg litre⁻¹ nitrite completely inhibited the growth of E coli O157: H7. While at higher pH levels, 2 g litre⁻¹ sorbate or 1 g litre⁻¹, nitrite, the highest concentration tested, did not show significant antimicrobial action against the test organisms. At 4°C and pH 5·0, the inoculated test organisms did not showed any significant growth in preservative-free control media. Different degree of inactivation and injury was observed when E coli O157: H7 strain 933 was stored in TSB (pH 5·0) containing 1 g litre⁻¹ sorbate or nitrite at 37°C. At 4°C, inactivation and injury of E coli O157: H7 cells was not observed in the medium containing sorbate or nitrite throughout the 24 h experimental period.     

大肠杆菌O157:H7快速培养的初步研究

本研究将Oxyrase酶加入到接种了大肠杆菌O157:H7的培养基中以促进这种兼性厌氧微生物的生长。与不加Oxyrase酶的对照组相比,添加了Oxyrase酶的培养基中的大肠杆菌O157:H7的浓度明显提高。实验结果表明,Oxyrase酶在大肠杆菌O157:H7 快速培养中具有潜在的利用价值。     

抗大肠杆菌O157:H7的卵黄特异性IgY的研究

以大肠杆菌O157:H7为抗原免疫产蛋母鸡,从鸡卵黄中提取免疫球蛋白,建立抗大肠杆菌O157:H7的特异性IgY的效价检测方法,并研究母鸡的免疫应答性,以及抗体的提取方法和体外抑菌效果.研究结果表明,初次免疫后第6d,在卵黄中可以检测到抗大肠杆菌O157:H7 IgY,效价为1:7200;经加强免疫后效价迅速上升,至第44d达到最高效价1:230400;免疫后360 d,效价仍维持在1:7200.用水稀释法、硫酸铵分级盐析和Sephadex G-25凝胶过滤以提取IgY,提纯后IgY的效价是之前的4倍.SDS-PAGE鉴定抗体的纯度,电泳图谱中出现抗体的轻链和重链两条带.体外抑菌实验表明,IgY能抑制大肠杆菌O157:H7的生长.     

Effect of Meat Binding Formulations on Thermal Inactivation of Escherichia coli O157:H7 Internalized in Beef

ABSTRACT:  This study evaluated the effects of meat binding or restructuring formulations, including salt/phosphate, algin/calcium, Activa™RM, and Fibrimex®, with or without 0.27% (wt/wt) lactic acid, on thermal inactivation of internalized Escherichia coli O157:H7 in ground beef, serving as a model system for restructured products. Ground beef batches (700 g; approximately 5% fat) were mechanically mixed with a 5-strain composite of E. coli O157:H7 (7 log CFU/g) and then with the restructuring formulations. Product portions (30 g) were extruded into plastic test tubes (2.5 × 10 cm) and stored at 4 °C (18 h), before heating to 60 or 65 °C in a circulating water bath to simulate rare or medium-rare doneness of beef, respectively. Cooking to 60 or 65 °C reduced ( P < 0.05) bacterial counts of control samples by 1.8 and 3.2 log CFU/g, respectively. Thermal destruction at 60 °C was not different ( P > 0.05) among all treatments and the control. At 65 °C, greater ( P < 0.05) thermal inactivation of E. coli O157:H7, as compared to the control, was obtained in samples treated with lactic acid alone (reductions of 4.9 log CFU/g), whereas for all other treatments, microbial destruction (reductions of 2.2 to 4.5 log CFU/g) was comparable ( P > 0.05) to that of the control. Cooking weight losses were lower ( P < 0.05) in salt/phosphate samples (<1%) compared to other formulations and the control (7.4% to 15.9%). Findings indicated that, under the conditions examined, restructuring of beef with salt/phosphate, algin/calcium, Activa™RM, or Fibrimex® did not affect inactivation of internalized E . coli O157:H7 in undercooked (60 or 65 °C) samples, whereas inclusion of lactic acid (0.27%) in nonintact beef products enhanced pathogen destruction at 65 °C.     

Cold plasma reduction of Salmonella and Escherichia coli O157:H7 on almonds using ambient pressure gases

Contamination of raw nuts, including almonds, is a food safety concern. Cold plasma is a novel antimicrobial intervention that can eliminate foodborne pathogens. The objective of this work was to evaluate the efficacy of rapid cold plasma treatments in eliminating Salmonella and Escherichia coli O157:H7 from dry almonds. Three isolates of Salmonella (S. Anatum F4317, S. Stanley H0558, and S. Enteritidis PT30) and 3 isolates of E. coli O157:H7 (C9490, ATCC 35150, and ATCC 43894) were separately grown and spot-inoculated (10 μL) onto whole almonds and allowed to dry for 10 min. Inoculated almonds were treated with a cold plasma jet, with treatment variables evaluated in a factorial design for each isolate: time, distance, and feed gas. Treatment time was 0 s (control), 10 s, or 20 s. Distance from the emitter was 2, 4, or 6 cm. Feed gas was dry air or nitrogen. After treatment, the almonds were sampled using swabs. Survivors were enumerated on tryptic soy agar (TSA) plates. Cold plasma significantly reduced both pathogens on almonds. The greatest reduction observed was 1.34 log cfu/mL reduction of E. coli O157:H7 C9490 after 20 s treatment at 6 cm spacing. The interaction of treatment time with distance from plasma emitter head was complex, and isolate-dependent. Longer duration of treatment did not always result in enhanced reductions. In general, nitrogen as a feed gas resulted in a reduced antimicrobial efficacy compared to dry air. These results indicate that short pulses of atmospheric pressure cold plasma can significantly reduce Salmonella and E. coli O157:H7 on almonds.     

Antimicrobial Effects of Quillaja saponaria Extract Against Escherichia coli O157:H7 and the Emerging Non‐O157 Shiga Toxin‐Producing E. coli

Natural alternate methods to control the spread of Shiga toxin‐producing Escherichia coli (STEC) are important to prevent foodborne outbreaks. Quillaja saponaria aqueous bark extracts (QE), cleared by the U.S. Food and Drug Administration as a natural flavorant, contain bioactive polyphenols, tannins, and tri‐terpenoid saponins with anti‐inflammatory and antimicrobial activity. The objective of this study was to determine the effects of commercial QE against E. coli O157:H7 and non‐O157 strains over 16 h at 37 °C and RT. Overnight cultures of 4 E. coli O157:H7 strains and 6 non‐O157 STECs in Tryptic Soy Broth (TSB) were washed and resuspended in phosphate‐buffered saline (PBS, pH 7.2), and treated with QE and controls including citric acid (pH 3.75), sodium benzoate (0.1% w/w), acidified sodium benzoate (pH 3.75) or PBS for 6 h or 16 h. Recovered bacteria were enumerated after plating on Tryptic Soy Agar, from duplicate treatments, replicated thrice and the data were statistically analyzed. The 4 QE‐treated E. coli O157:H7 strains from initial ～7.5 log CFU had remaining counts between 6.79 and 3.5 log CFU after 16 h at RT. QE‐treated non‐O157 STECs showed lower reductions with remaining counts ranging from 6.81 to 4.55 log CFU after 16 h at RT.  Incubation at 37 °C caused reduction to nondetectable levels within 1 h, without any significant reduction in controls. Scanning electron microscopy studies revealed damaged cell membranes of treated bacteria after 1 h at 37 °C. QE shows potential to control the spread of STECs, and further research in model food systems is needed.     

应用DPO-PCR技术检测肠出血性大肠杆菌O157∶H7

利用双启动寡核苷酸引物（dual-priming oligonucleotide,DPO）聚合酶链式反应（polymerase chain reaction,PCR）技术检测肠出血性大肠杆菌O157∶H7。根据DPO引物设计原则,以肠出血性大肠杆菌O157∶H7 rfbE 基因为靶基因设计一对DPO引物,经过反应体系的优化,建立了肠出血性大肠杆菌O157∶H7 DPO-PCR检测方法, 其检测灵敏度约为94 CFU/mL。与常规PCR方法相比,所建立的DPO-PCR方法对退火温度不敏感,在引物设计和实 验过程中不需要对引物及其退火温度反复优化,同时基于DPO引物的特殊结构又增强了其特异性。DPO-PCR方法 设计简易、特异性强,为致病性微生物的快速准确检测提供了新途径。