In vinegar production, oxygen has to be made available for the production of acetic acid, whereas with wine it is essential to exclude oxygen to prevent oxidation of the alcohol and spoilage of the wine. 0000018861 00000 n While the total bacterial number in an oil or gas production sample indicates the general conditions for bacteria to grow in this unique environment, and, thus, the potential risk of microbial corrosion, some specific groups of microorganisms commonly found in such samples probably play a more important role in the corrosion process. �*T0T0 Bi������f����� � In addition, all primer sequences were analyzed for specificity using a BLAST SEARCH FOR SHORT, NEARLY EXACT MATCHES program and were found not to cross-react with any other non-target organisms. 0000002408 00000 n <>>>/Contents 21 0 R/Parent 3 0 R>> The gene ackA encodes the key enzyme acetate kinase α subunit, which along with phosphotransacetylase, catalyzes the conversion of acetyl coenzyme A to acetate. A qPCR was performed using the primers of this invention, a Rotor-Gene 3000, 4 Channel Multiplexing, real-time PCR system (Corbett Research, Sydney, Australia), and a QuantiTect SYBR Green PCR kit (Qiagen, Inc., Valencia, Calif.). <> <>>>/Contents 9 0 R/Parent 3 0 R>> MeltCal software, version 2.1 was then used to check the cross-hybridization among the sequences of primers used in some assay and calculate the melting temperature (Tm) using a nearest neighbor model. endobj Quantitative PCR has also been used to determine the abundance of microorganisms in many different types of complex environmental samples such as sediments, water, wastewater, and marine samples. 0000015682 00000 n aureus N315 (BA000018), and Staphylococcus aureus subsp. Microbiologically influenced corrosion is a significant problem affecting the gas and oil industry as well as other industries, and the monitoring of microbial populations within such industries is a component of overall corrosion control operations. 24 0 obj <>/ProcSet[/PDF/Text/ImageB]/XObject<>>>/Type/Page>> endobj %���� [55 0 R] 0000001203 00000 n To address the above-stated object as well as other objects of this invention, a real-time qPCR assay has been developed by targeting the gene ackA encoding acetate kinase α subunits in acetic acid-producing bacteria. Several species of acetic acid bacteria are used in industry for production of certain foods and chemicals. AAB of the family Acetobacteraceae are ubiquitous and are known to be adapted to various sugar- and ethanol-rich environments. The primers are particularly suitable for amplifying the nucleic acid of acetic acid-producing bacteria present in gas and oil production operations. <>>>/Contents 12 0 R/Parent 3 0 R>> All of these bacteria have the ackA gene. 0000046963 00000 n 0000002158 00000 n Acetic acid bacteria are essential for brewing various vinegars. The acetic acid bacteria consist of 10 genera in the family Acetobacteraceae. endobj 12 0 obj trailer However, cycles numbers outside of this range may be necessary and, thus, the statement of the above stated range is in no way intended to limit the scope of this invention. Microbiol., 2004. Thus, the PCR primer pairs are ideally suited to accurately quantify acetic acid-producing bacteria in gas and oil production operations that have relevance to microbiologically influenced corrosion. This invention relates to a method for the rapid detection and quantification of acetic acid-producing bacteria using real-time quantitative polymerase chain reactions (qPCR). Gas Technology Institute (Des Plaines, IL, US). SYBR Green I, in a solution for binding therebetween, and the acetic acid-producing bacteria is quantified by monitoring the signal produced by the bound label, the intensity of which is a function of the quantity of the ackA gene in acetic acid-producing bacteria in the sample. 0000026466 00000 n DNA amplification reactions include polymerase chain reactions, PCR, which may consist of 10 to 50 cycles of denaturization and synthesis of a DNA molecule. As used herein, the term “amplification” refers to any in vitro method for increasing the number of copies of a nucleotide sequence with the use of a DNA polymerase. %PDF-1.7 %���� <>/ProcSet[/PDF/Text/ImageB]/XObject<>>>/Type/Page>> endobj 1-47, 2005. %%EOF If fermentation occurs in a medium rich in glutamic acid, this acid could lead to the formation of succinic acid. Acetic acid bacteria (AAB) are a group of Gram-negative bacteria which oxidize sugars or ethanol and produce acetic acid during fermentation. 120 of samples were collected, including 40 raw milk, 40 cheeses and 40 yoghurt were collected randomly from Babylon province. The cycling conditions consisted of 15 minutes of incubation at 95° C. followed by 35 cycles of 95° C. for 30 sec, 52° C. for 30 sec, and 73° C. for 30 sec for denaturing, annealing and elongation, respectively, followed by 70° C. for 30 sec, and 70° C. to 99° C. to generate melting curves. �26S073QI�r � 19 0 obj 2. ��Y�"�2Y�͕�*���;���=��cؠ �n�5�Li��AQ��� �}�?�'��7�=������-. ?~\��ǜ����v��e�2�j�~Jې@���/��B���4ϻ$�jb�p��$�I�*���{���ω6�64׈`���X3��FG�=�;��Ih4-�R�3�"]&��e^K����d/��Ρ�Ty)����fԗ��8VG�d�-\�a{h������L�X��f�){.�\�1>��SI�2W�@�c���Մ��$�¨JaT�E^��NB �-��& ����,�l#��`�'��J˼4��~$~�oX�����w�Uz� �*T0T0 Bi������f����� � Journal of American Sciencew, vol. A primer, complementary to a portion of a template is hybridized under appropriate conditions and a polymerase then synthesizes a molecule complementary to the template or a portion thereof.