Species specific identification of spore-producing microbes using the gene sequence of small acid-soluble spore coat proteins for amplification based diagnostics
AssigneeThe Regents of the University of California
InventorNancy McKinney
Life of this patent
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Abstract
PCR (polymerase chain reaction) primers for the detection of certain Bacillus species, such as Bacillus anthracis. The primers specifically amplify only DNA found in the target species and can distinguish closely related species. Species-specific PCR primers for Bacillus anthracis, Bacillus globigii and Clostridium perfringens are disclosed. The primers are directed to unique sequences within sasp (small acid soluble protein) genes.
Claims15
A PCR primer useful in detecting Bacillus anthracis, comprising the sequence:
- 5′ GTT ATG GTA CAG AGT TTG CG 3′ (SEQ ID NO. 94).
A PCR primer useful in detecting Bacillus anthracis, comprising the sequence:
- 5′ TTG TTT TGC AGC TGA TTG T 3′ (SEQ ID NO. 95).
A method for specifically detecting a DNA sequence from Bacillus anthracis comprising the steps of:
- (a) providing a DNA sample;
- (b) incubating the DNA sample with primers comprising the sequences:
- 5′ GTT ATG GTA CAG AGT TTG CG 3′ (SEQ D NO. 94) and
- 5′ TTG TTT TGC AGC TGA TTG T 3′ (SEQ D NO. 95);
- (c) amplifying DNA hybridizing to said primers;
- and
- (d) detecting the presence of any amplified product,
- whereby only DNA from Bacillus anthracis is amplified and detected.
A PCR primer useful in detecting Bacillus globigii, consisting of the sequence:
- 5′ ACA TGG CTA ACT CAA ACA ACA A 3′ (SEQ ID NO. 103).
A PCR primer useful in detecting Bacillus globigii, comprising the sequence:
- 5′ GGT TTT GTT TTC TTA CTT GTT GTA C 3′ (SEQ ID NO 104).
A method for specifically detecting a DNA sequence from Bacillus globigii, comprising the steps of:
- (a) providing a DNA sample;
- (b) incubating the DNA sample with primers comprising the sequences:
- 5′ ACA TGG CTA ACT CAA ACA ACA A 3′ (SEQ ID NO. 103) and
- 5′ GGT TTT GTT TTC TTA CTT GTT GTA C 3′ (SEQ ID NO. 104);
- (c) amplifying DNA hybridizing to said primers;
- and
- (d) detecting the presence of any amplified product,
- whereby only DNA from Bacillus globigii is amplified and detected.
A PCR primer useful in detecting Clostridium perfringens, consisting essentially of the sequence:
- 5′ AAT AAC TAA GGA GGA ATG AAA AAT GT 3′ (SEQ ID NO. 105).
A PCR primer useful in detecting Clostridium perfringens, consisting essentially of the sequence:
- 5′ TTG TTC TAC CAT TCT TTT AAC CAT T 3′ (SEQ ID NO. 106).
A method for specifically detecting a DNA sequence from Clostridium perfringens, comprising the steps of:
- (a) providing a DNA sample;
- (b) incubating the DNA sample with primers consisting essentially of the sequences:
- 5′ AAT AAC TAA GGA GGA ATG AAA AAT GT 3′ (SEQ ID NO.105) and
- 5′ TTG TTC TAC CAT TCT TTT AAC CATT 3′ (SEQ ID NO. 106);
- (c) amplifying DNA hybridizing to said primers;
- and
- (d) detecting the presence of any amplified product,
- whereby DNA from Clostridium perfringens is amplified and detected.
A cloned and isolated nucleic acid encoding a Bacillus globigii sasp-gamma polypeptide comprising the sequence recited in SEQ ID NO 110.
A method for the detection of Bacillus globigii in a sample, comprising the steps of:
- (a) specifically incubating the sample with amplification primers that hybridize to the Bacillus globigii sasp-gamma gene;
- (b) amplifying hybridized primers;
- and
- (c) detecting the presence of amplified Bacillus globigii sasp-gamma gene sequences.
A primer which hybridizes completely to the sequence SEQ ID NO:
- 110 and which is capable of amplifying specifically SEQ ID NO:
- 110.
Dependencies are inferred from the claim text.