公开(公告)号：US20090203017A1

公开(公告)日：2009-08-13

申请号：US12359263

申请日：2009-01-23

Applicant: ELLIOTT P. DAWSON ,  STEVEN J. SIMMONS ,  LORI J. RAY-COX ,  JENNIFER M. BAKER ,  KRISTIE E. WOMBLE ,  JUDITH MADDEN ,  SUBRAMANI SELLAPPAN ,  ANDREW HEARN ,  SAL SEMINARA

Inventor： ELLIOTT P. DAWSON ,  STEVEN J. SIMMONS ,  LORI J. RAY-COX ,  JENNIFER M. BAKER ,  KRISTIE E. WOMBLE ,  JUDITH MADDEN ,  SUBRAMANI SELLAPPAN ,  ANDREW HEARN ,  SAL SEMINARA

IPC: C12Q1/68

CPC classification number: C12Q1/6816 ,  C12Q2563/107 ,  C12Q2521/301

Abstract: The invention relates to methods for the determination and detection of nucleic acids sequences in a sample. The nucleic acid may be RNA or DNA or both. The invention also relates to methods for the determination of the presence and species of various microorganisms in a sample. We have also identified a set of oligonucleotide nucleic acid sequences within the rRNAs of Gram-negative organisms that facilitates both the broad identification of Gram-negative organisms as a class when used as a pool, or in combination, for example in a hybridization assay. This set of oligonucleotides may detect sequences that are indicative of the presence of organisms of the broad class of Gram-negative organisms while exhibiting little or no false identification of Gram-positive organisms, and fungi, or other microorganisms. The assay includes concurrent incubation with at least one nucleotide sequence of interest, at least one nucleic acid probe, a fluorosurfactant, and a nuclease. The assay may further be employed to detect the presence of bacteria, fungi, or other microorganisms by use of additional specific probes, or to detect and/or identify target nucleic acid sequences in a sample. Further, the invention also relates to methods of reducing non-specific binding and facilitating complex formation in a binding assay. The binding assay may be, but is not limited to, a nucleic acid hybridization assay or an immunoassay. The invention also relates to methods of detection that employ at least one target of interest, which may be a nucleotide sequence, at least one probe, which may be a nucleic acid probe and a nuclease.

Abstract translation: 本发明涉及用于测定和检测样品中核酸序列的方法。 核酸可以是RNA或DNA或两者。 本发明还涉及用于确定样品中各种微生物的存在和种类的方法。 我们还在革兰氏阴性生物体的rRNAs中鉴定了一组寡核苷酸核酸序列，当用作池或组合使用时，例如在杂交测定中，可以广泛鉴定革兰氏阴性生物体。 这组寡核苷酸可以检测指示广泛类型的革兰氏阴性生物体的生物体的存在的序列，同时表现出对革兰氏阳性生物体，真菌或其它微生物的很少或没有虚假的鉴定。 该测定包括与至少一个目的核苷酸序列，至少一个核酸探针，含氟表面活性剂和核酸酶的同时孵育。 该测定法还可用于通过使用另外的特异性探针来检测细菌，真菌或其它微生物的存在，或检测和/或鉴定样品中的靶核酸序列。 此外，本发明还涉及减少非特异性结合并促进结合测定中复合物形成的方法。 结合测定可以是但不限于核酸杂交测定或免疫测定。 本发明还涉及使用至少一种感兴趣靶标（其可以是核苷酸序列）的至少一种探针，其可以是核酸探针和核酸酶的检测方法。

公开(公告)号：US20120301907A1

公开(公告)日：2012-11-29

申请号：US13537267

申请日：2012-06-29

Applicant: SUBRAMANI SELLAPPAN ,  ANDREW HEARN ,  SALVATORE SEMINARA

Inventor： SUBRAMANI SELLAPPAN ,  ANDREW HEARN ,  SALVATORE SEMINARA

IPC: G01N21/76 ,  C12M1/34 ,  G01N1/34

CPC classification number: C12Q1/04 ,  G01N1/38 ,  G01N21/76 ,  G01N21/77 ,  G01N33/56911 ,  G01N2001/028 ,  G01N2001/4061 ,  G01N2001/4088 ,  G01N2201/02

Abstract: A rapid, sensitive method of separating and detecting microorganisms from a sample potentially containing microorganisms, such as but not limited to bacteria, fungi, yeast, viruses, and the like. The method relies on separation techniques to separate and concentrate the cells from the sample, together with chemical techniques to amplify the amount of detectable signal from low numbers of cells to provide a rapid and sensitive method of detecting microorganisms. This detection method may utilize: a filtration device; a centrifugation device; a system; a swab device; and kit comprising one or more of the devices and components to perform the present method of separating and detecting microorganisms in a sample potentially containing microorganisms. The sample may be a chemical, cosmetic, personal care, pharmaceutical, or consumable good in its raw material, in-process, and/or finished product states that needs to be tested for any contaminating microorganisms prior to shipment to the consumer.

Abstract translation: 从可能含有微生物（例如但不限于细菌，真菌，酵母，病毒等）的样品中分离和检测微生物的快速，灵敏的方法。 该方法依赖于分离技术以将样品中的细胞与化学技术一起分离和浓缩，从而扩增来自低细胞数量的可检测信号的量，以提供检测微生物的快速和灵敏的方法。 该检测方法可以利用：过滤装置; 离心装置; 一个系统; 拭子装置 以及包含一个或多个装置和组件的试剂盒，以执行分离和检测可能含有微生物的样品中的微生物的本方法。 样品可能是化学，化妆品，个人护理，药物或消耗品的原材料，在制品和/或成品状态，需要在装运到消费者之前对任何污染的微生物进行测试。

公开(公告)号：US20080182235A1

公开(公告)日：2008-07-31

申请号：US11669078

申请日：2007-01-30

Applicant: Andrew Hearn ,  Judith Madden ,  Subramani Sellappan ,  Antje J. Baeumner ,  Natalya V. Zaytseva

Inventor： Andrew Hearn ,  Judith Madden ,  Subramani Sellappan ,  Antje J. Baeumner ,  Natalya V. Zaytseva

IPC: C12Q1/70 ,  C12Q1/48 ,  G01N33/53 ,  G01N33/554 ,  C12Q1/68

CPC classification number: G01N33/5432

Abstract: The invention relates to the encapsulation of luminescence-related molecules, including but not limited to, adenosine triphosphate (ATP), adenylate kinase (AK), alkaline phosphatase (ALP), luminol and luciferin/luciferase cocktails, within liposomes. These liposomes can be employed to enhance the luminescence detection of microorganisms and compounds in various products and samples. The liposomes containing the luminescence-related molecules can bear a probe which has a specific sequence or structure that, in turn can be used to hybridize to, or couple with, a portion of the target analyte. Within the same assay, paramagnetic beads can bear a probe having a specific sequence or structure that, can hybridize to, or couple with, a second portion of the target analyte to create a complex of analyte bound to paramagnetic beads and liposomes. This type of assay can be often referred to as a ‘sandwich’ assay. Once the probes hybridize to, or couple with, their targets, a complex can be formed of the paramagnetic beads, the analyte, or portion thereof, and the liposomes. This complex can then be washed to remove those components that are non-hybridized or non-coupled. Then, the paramagnetic bead-analyte-liposome complexes can be isolated from the sample using magnetic separation techniques and can be treated so as to release their encapsulated ATP, AK or other luminescence-related compounds. The resulting luminescence can then be determined in a chemical assay. This determination can be qualitative (i.e., an absence/presence assay) or quantitative (i.e., which can measure a specific amount of analyte present). Through the use of a cocktail of probe types, the assay can also qualitatively or quantitatively measure the presence of more than one analyte simultaneously. This type of assay can be of commercial importance in clinical and forensic applications, the personal care, pharmaceutical, food and beverage markets, as well as in environmental sample assays.

公开(公告)号：US09932620B2

公开(公告)日：2018-04-03

申请号：US13537267

申请日：2012-06-29

Applicant: Subramani Sellappan ,  Andrew Hearn ,  Salvatore Seminara

Inventor： Subramani Sellappan ,  Andrew Hearn ,  Salvatore Seminara

IPC: C12M1/34 ,  G01N21/76 ,  G01N1/34 ,  C12Q1/04 ,  G01N21/77 ,  G01N1/38 ,  G01N33/569 ,  G01N1/02 ,  G01N1/40

CPC classification number: C12Q1/04 ,  G01N1/38 ,  G01N21/76 ,  G01N21/77 ,  G01N33/56911 ,  G01N2001/028 ,  G01N2001/4061 ,  G01N2001/4088 ,  G01N2201/02

Abstract: A rapid, sensitive method of separating and detecting microorganisms from a sample potentially containing microorganisms, such as but not limited to bacteria, fungi, yeast, viruses, and the like. The method relies on separation techniques to separate and concentrate the cells from the sample, together with chemical techniques to amplify the amount of detectable signal from low numbers of cells to provide a rapid and sensitive method of detecting microorganisms. This detection method may utilize: a filtration device; a centrifugation device; a system; a swab device; and kit comprising one or more of the devices and components to perform the present method of separating and detecting microorganisms in a sample potentially containing microorganisms. The sample may be a chemical, cosmetic, personal care, pharmaceutical, or consumable good in its raw material, in-process, and/or finished product states that needs to be tested for any contaminating microorganisms prior to shipment to the consumer.