WO2022018685 COMPOSITIONS AND METHODS FOR ONE-STEP NUCLEIC ACID AMPLIFICATION AND DIAGNOSTIC METHODS BASED THEREON

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Publication Number WO/2022/018685

Publication Date 27.01.2022

International Application No. PCT/IB2021/056646

International Filing Date 22.07.2021

IPC

CPC

Applicants

KING ABDULLAH UNIVERSITY OF SCIENCE AND TECHNOLOGY [SA]/[SA]

Inventors

HAMDAN, Samir M.
TAKAHASHI, Masateru
TEHSEEN, Muhammad
TAKAHASHI, Etsuko

Priority Data

63/055,250		22.07.2020		US
63/214,738		24.06.2021		US

Publication Language English (en)

Filing Language English (EN)

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Title

(EN) COMPOSITIONS AND METHODS FOR ONE-STEP NUCLEIC ACID AMPLIFICATION AND DIAGNOSTIC METHODS BASED THEREON

(FR) COMPOSITIONS ET PROCÉDÉS D'AMPLIFICATION D'ACIDE NUCLÉIQUE EN UNE ÉTAPE ET PROCÉDÉS DE DIAGNOSTIC BASÉS SUR CELLES-CI

Abstract

(EN) Compositions and methods for a nucleic acid amplification and diagnostic methods based therein, are disclosed. The compositions include purified Thermus aquaticus DNA polymerase (Taq Pol) and Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV-RT), in proportions and under reaction conditions that allow one-step amplification of a nucleic acid of interest. In particular, the compositions are useful in one-step RT-qPCR and RT-LAMP methods. The method includes sampling the specimen from a subject, extracting and purifying RNA from the sample. The compositions and methods may be used to produce, analyze, quantitate, detect and otherwise manipulate nucleic acid molecules. For example, disclosed are methods of detecting the presence of a viral nucleic acid in a sample from a subject. Methods of diagnosing a subject as being infected with a viral pathogen are also provided. A preferred virus or pathogen is SARS-CoV-2.

(FR) L'invention concerne des compositions et des procédés pour une amplification d'acide nucléique et des procédés de diagnostic basés sur celles-ci. Les compositions comprennent de l'ADN polymérase de Thermus aquaticus (Taq Pol) et transcriptase inverse du virus de la leucémie de Moloney Murine (MMLV-RT) purifiées, dans des proportions et des conditions de réaction qui permettent l'amplification en une étape d'un acide nucléique d'intérêt. En particulier, les compositions sont utiles dans les procédés RT-qPCR et RT-LAMP en une étape. Le procédé comprend l'échantillonnage de l'échantillon prélevé sur un sujet, l'extraction et la purification de l'ARN à partir de l'échantillon. Les compositions et les procédés peuvent être utilisés pour produire, analyser, quantifier, détecter et manipuler autrement des molécules d'acide nucléique. Par exemple, l'invention concerne des procédés de détection de la présence d'un acide nucléique viral dans un échantillon provenant d'un sujet. L'invention concerne également des procédés de diagnostic d'un sujet comme étant infecté par un pathogène viral. Un virus ou agent pathogène préféré est le SARS-CoV-2.

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q	MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1	Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
70	involving virus or bacteriophage

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q	MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1	Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68	involving nucleic acids
6844	Nucleic acid amplification reactions
686	Polymerase chain reaction

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q	MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1	Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
70	involving virus or bacteriophage

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q	MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1	Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68	involving nucleic acids
6844	Nucleic acid amplification reactions
686	Polymerase chain reaction

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q	MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1	Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68	involving nucleic acids
6844	Nucleic acid amplification reactions
686	Polymerase chain reaction [PCR]

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1. WO2022018685 - COMPOSITIONS AND METHODS FOR ONE-STEP NUCLEIC ACID AMPLIFICATION AND DIAGNOSTIC METHODS BASED THEREON