1. A molecular-beacon-based multi-allelic real-time reverse transcription polymerase chain reaction (RT-PCR) multiplex assay for a Sever Acute Respiratory Syndrome (SARS) virus comprising:
(1) obtaining a sample;
(2) isolating RNA from the sample;
(3) placing the isolated RNA in a tube along with primers specific for spike (S), envelope (E), membrane (M), and nucleocapsid (N) genes of the SARS virus, wherein the S primer includes SEQ ID NO:18, the E primer includes SEQ ID NO:38, the M primer includes SEQ ID NO:59, and the N primer includes SEQ ID NO:80,
(4) reverse transcribing the isolated RNA in the presence of a reverse transcriptase to form a cDNA unique for each of the S, E, M and N SARS viral genes,
(5) real-time amplifying the SARS cDNA in the presence of four distinct types of molecular beacons, each labeled with the same fluorophore and specific for a different SARS coronavirus (CoV) gene selected from S, E, M and N viral genes, and four S, E, M and N gene-specific primer pairs,
wherein the molecular beacons are:
for S gene:
LK249 5\u2032-FAM-CCCACGCCAGAAGGTAGATCACGAACTACACGTGGG-3\u2032-Dubcyl (SEQ ID NO:14);
for E gene:
LK253 5\u2032-FAM-CCTCCGCACGAAAGCAAGAAAAAGAAGTACGCCGGAGG-3\u2032-Dubcyl (SEQ ID NO:34);
for M gene:
LK257 5\u2032-FAM-CCTCCGACCCAATTAATTCTGTAGACAGCAGCCGGAGG-3\u2032-Dubcyl (SEQ ID NO:55); and
for N gene:
LK261 5\u2032-FAM-CCTCCGTACCATCTGGGGCTGAGCTCTTFCATCGGAGG-3\u2032-Dubcyl (SEQ ID NO:76);
wherein the S, E, M and N gene-specific primer pairs are:
for S gene
LK251 5\u2032-CTCTATGTTTATAAGGGCTATCAACC-3\u2032 (SEQ ID NO:16)
LK252 5\u2032-CCAAGAGGCAACTTAAAAATAGGTTTC-3\u2032 (SEQ ID NO:17);
for E gene
LK255 5\u2032-CGGAAGAAACAGGTACGTTAATAG-3\u2032 (SEQ ID NO:36)
LK256 5\u2032-AAGCGCAGTAAGGATGGCTA-3\u2032 (SEQ ID NO:37);
for M gene
LK259 5\u2032-CTTGTTTTCCTCTGGCTCTTG-3\u2032 (SEQ ID NO:57)
LK260 5\u2032-CAAGCCATTGCAATCGCAATC-3\u2032 (SEQ ID NO:58); and
for N gene
LK263 5\u2032-ACGAGTTCGTGGTGGTGAC-3\u2032 (SEQ ID NO:78)
LK264 5\u2032-CGTAGGGAAGTGAAGCTTC-3\u2032 (SEQ ID NO:79); and
(6) measuring fluorescence, which is a collective measure of the presence of the SARS virus.
2. The assay of claim 1 wherein the SARS virus is SARS-CoV.
3. The assay of claim 1, wherein step (3) further comprises a RNA internal positive control (IPC) and step (5) further comprises IPC-specific primers and a fifth molecular beacon that is specific for the IPC and labeled with a different fluorophore, which can be distinguished from the SARS-CoV gene-specific molecular beacon fluorophore, and wherein step (6) further includes a measurement of fluorescence at a wavelength unique for the fifth molecular beacon.
4. The assay of claim 1 wherein the sample source includes blood, tissue, sputum, or nasopharyngeal aspirations.
5. The assay of claim 1 wherein the measuring step includes simultaneous automated amplicon detection in a spectrofluorometric thermal cycler that measures the fluorescence intensity of each color during the annealing phase of each thermal cycle.
6. The assay of claim 3 wherein the IPC-specific primers are included in step (5) and have the following sequences: SEQ ID NO:16 and SEQ ID NO:37.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A voice encoder comprising:
a quantization selection unit generating a quantization selection signal to represent a result of a selecting, before quantizing a line spectral frequency (LSF) of a current frame of an input signal, one of a first LSF quantization unit and a second LSF quantization unit for the quantizing of the LSF of the current frame, wherein the selecting is based on analysis by the quantization selection unit of a generated synthesized voice signal of a previous frame of the input signal; and
a quantization unit extracting a linear prediction coding (LPC) coefficient from the current frame of the input signal, converting the extracted LPC coefficient into the LSF of the current frame, quantizing the LSF of the current frame with the selected one of the first LSF quantization unit using a first predictor and the second LSF quantization unit using a second predictor, the second predictor being different from the first predictor, based on the quantization selection signal, and converting the quantized LSF into a quantized LPC coefficient.
2. The voice encoder according to claim 1, wherein the quantization unit includes:
an LPC coefficient extraction unit to extract a LPC coefficient of the previous frame from the input signal;
an LSF conversion unit to convert the extracted LPC coefficient of the previous frame into an LSF of the previous frame;
the first LSF quantization unit to quantize the LSF of the previous frame through a first quantization process;
the second LSF quantization unit to quantize the LSF of the previous frame through a second quantization process; and
an LPC coefficient conversion unit to convert a quantized LSF of the previous frame, generated by a selected one of the first LSF quantization unit and the second LSF quantization unit to perform quantizing of the LSF of the previous frame, into a quantized LPC coefficient of the previous frame.
3. The voice encoder according to claim 2, wherein the LPC quantization unit extracts the LPC coefficient corresponding to the current frame using autocorrelation and a Levinson-Durbin algorithm.
4. The voice encoder according to claim 2, wherein the LSF conversion unit outputs the LSF of the previous frame to a selected one of the first quantization unit and the second LSF quantization unit according to a quantization selection signal generated for the selecting of the first LSF quantization unit and the second LSF quantization unit one of the quantizing of the LSF of the frame.
5. The voice encoder according to claim 1, wherein the quantization selection unit includes:
an energy variation calculation unit to calculate energy variations of the synthesized voice signal of at least the previous frame;
a zero crossing calculation unit to calculate a changing degree of a sign of the synthesized voice signal of at least the previous frame;
a pitch difference calculation unit to calculate a pitch delay of the synthesized voice signal of at least the previous frame; and
a selection signal generation unit checking whether the synthesized voice signal of at least the previous frame has a voice signal based on the calculated energy variation, and generating the quantization selection signal based on a result of the checking indicating that the synthesized voice signal of at least the previous frame has the voice signal, the calculated changing degree of the sign of the synthesized voice signal of at least the previous frame, and the calculated pitch delay of the synthesized voice signal of at least the previous frame.
6. The voice encoder according to claim 5, wherein the energy variation calculation unit includes:
an energy calculation unit to calculate energy values in respective subframes constituting at least the previous frame;
an energy buffer to store the calculated energy values of the respective subframes;
a moving average calculation unit to calculate a moving average for the stored energy values of the respective subframes; and
an energy increasedecrease calculation unit to calculate energy variation in at least the previous frame based on the calculated moving average and the calculated energy values of the respective subframes.
7. The voice encoder according to claim 1, further comprising:
a perceptual weighting filter perceptually weighting the input signal based on a quantized LPC coefficient of the previous frame;
a subtractor subtracting a specified synthesized signal from the perceptually weighted input signal to generate a linear prediction remaining signal; and
a signal synthesis unit searching for an excited signal from the linear prediction remaining signal, generating the specified synthesized signal using the quantized LPC coefficient of the previous frame and an excited signal found in the searching, and outputting the specified generated synthesized signal to the subtractor.
8. A voice encoder comprising:
a quantization selection unit generating a quantization selection signal;
a quantization unit extracting a linear prediction coding (LPC) coefficient from a current frame of an input signal, converting the extracted LPC coefficient into a line spectral frequency (LSF), selectively quantizing the LSF with one of a first LSF quantization unit using a first predictor and a second LSF quantization unit using a second predictor, the second predictor being different from the first predictor, based on the quantization selection signal, and converting the quantized LSF into a quantized LPC coefficient of the current frame;
a perceptual weighting filter perceptually weighting the input signal based on a quantized LPC coefficient of a previous frame of the input signal;
a signal synthesis unit searching for an excited signal from a linear prediction remaining signal, generating a synthesized voice signal of the previous frame using the quantized LPC coefficient of the previous frame and an excited signal found in the searching, and outputting the generated synthesized voice signal to a subtractor;
the subtractor subtracting the synthesized voice signal from the perceptually weighted input signal to generate the linear prediction remaining signal; and,
wherein the quantization selection signal determines the selecting of the one of the first LSF quantization unit and the second LSF quantization unit based on characteristics of the synthesized voice signal, and
wherein the signal synthesis unit includes
a synthesis filter synthesizing the synthesized voice signal using a synthesized excited signal of the input signal, from an excited signal synthesis unit based on the found excited signal, and the quantized LPC coefficient of the previous frame, received from the LPC coefficient conversion unit, and outputting the synthesized voice signal to the subtractor and the quantization selection unit.
9. The voice encoder according to claim 8, wherein the linear prediction remaining signal is generated using the following equation:
x
\u2062
\u2062
(
n
)
=
s
w
\u2061
(
n
)
–
\u2211
i
=
1
10
\u2062
\u2062
a
^
i
\u2063
\xb7
s
^
\u2062
\u2062
(
n
–
i
)
\u2062
\u2062
n
=
0
,
\u2026
\u2062
,
L
–
1
wherein, x(n) is the linear prediction remaining signal, sw(n) is the perceptually weighted voice signal, \xe2i is an ith element of the quantized LPC coefficient vector, from the previous frame, \u015d(n) is the synthesized voice signal, and L is the number of sample per one frame.
10. A voice decoder comprising:
a dequantization selection unit generating a dequantization selection signal, the dequantization selection signal representing a result of a selecting, before dequantizing line spectral frequency (LSF) quantization information of a current frame of an input signal, one of a first LSF dequantization unit and a second LSF dequantization unit for the dequantizing of the LSF quantization information, wherein the selecting is based on analysis by the dequantization selection unit of a generated synthesized voice signal of a previous frame of the input signal; and
a dequantization unit dequantizing line spectral frequency (LSF) quantization information of the current frame to generate an LSF vector, and converting the LSF vector into a linear prediction coding (LPC) coefficient of the current frame, the LSF quantization information being received through a specified channel and dequantized using the selected one of the first LSF dequantization unit having a first predictor and the second LSF dequantization unit having a second predictor, the second predictor being different from the first predictor,
wherein the synthesized voice signal is generated from synthesis information of a received voice signal.
11. The voice decoder according to claim 10, wherein the dequantization unit includes:
the first LSF dequantization unit to generate an LSF vector of the previous frame through a first dequantization process of LSF dequantization information of the previous frame;
the second LSF dequantization unit to generate the LSF vector of the previous frame through a second dequantization process of the LSF dequantization information of the previous frame; and
an LPC coefficient conversion unit to convert the dequantized LSF vector of the previous frame, generated by a dequantizing of the LSF information using a selected one of the first LSF dequantization unit and the second LSF dequantization unit, into a dequantized LPC coefficient of the previous frame.
12. The voice decoder according to claim 10, wherein the dequantization selection unit includes:
an energy variation calculation unit to calculate energy variation of the synthesized voice signal of at least the previous frame;
a zero crossing calculation unit to calculate a changing degree of a sign of the synthesized voice signal of at least the previous frame;
a pitch difference calculation unit to calculate a pitch delay of the synthesized voice signal of at least the previous frame; and
a selection signal generation unit checking whether the synthesized voice signal of at least the previous frame has a voice signal based on the calculated energy variation, and generating a dequantization selection signal based on a result of the checking indicating that the synthesized voice signal of at least the previous frame has the voice signal, the calculated changing degree of the sign of the synthesized voice signal of at least the previous frame, and the calculated pitch delay of the synthesized voice signal of at least the previous frame.
13. The voice decoder according to claim 12, wherein the energy variation calculation unit includes:
an energy calculation unit to calculate energy values in respective subframes constituting at least the previous frame;
an energy buffer to store the calculated energy values of the respective subframes;
a moving average calculation unit to calculate a moving average for the stored energy values of the respective subframes; and
an energy increasedecrease calculation unit to calculate energy variation in at least the previous frame based on the calculated moving average and the calculated energy values of the respective subframes.
14. The voice decoder according to claim 11, further comprising a signal synthesis unit synthesizing an excited signal by using excited signal synthesis information of the input signal and the dequantized LPC coefficient of the previous frame received from the LPC coefficient conversion unit.
15. The voice decoder according to claim 14, further comprising an excited signal synthesis unit synthesizing the synthesize excited signal based on received excited signal synthesis information of the current frame, and outputting the synthesized excited signal to a synthesis filter filtering the synthesized excited signal.
16. The voice decoder according to claim 15, wherein the synthesized voice signal is synthesized according to the following equation:
s
^
\u2062
\u2062
(
n
)
=
x
^
\u2062
\u2062
(
n
)
+
\u2211
i
=
1
10
\u2062
\u2062
a
^
i
\u2063
\xb7
s
^
\u2062
\u2062
(
n
–
i
)
\u2062
\u2062
n
=
0
,
\u2026
\u2062
,
L
–
1
wherein {circumflex over (x)}(n) is the synthesized excited signal.
17. A method of selecting quantization in a voice encoder, the method comprising:
extracting a linear prediction encoding (LPC) coefficient from a current frame of an input signal;
converting the extracted LPC coefficient into a line spectral frequency (LSF) of the current frame;
generating a synthesized voice signal of a previous frame of the input signal;
selecting, before quantizing the LSF of the current frame, one of a first LSF quantization process and a second LSF quantization process for the quantizing of the LSF of the current frame, wherein the selecting is based on an analysis of the generated synthesized voice signal;
quantizing the LSF through the selected one of the first quantization process using a first predictor and the second LSF quantization process using a second predictor, the second predictor being different from the first predictor; and
converting the quantized LSF into an quantized LPC coefficient of the current frame.
18. A method of selecting quantization in a voice encoder, the method comprising:
extracting a linear prediction encoding (LPC) coefficient from an input signal;
converting the extracted LPC coefficient into a line spectral frequency (LSF);
selectively quantizing the LSF through one of a first quantization process using a first predictor and a second LSF quantization process using a second predictor, the second predictor being different from the first predictor, based on characteristics of a synthesized voice signal in previous frames of the input signal; and
converting the quantized LSF into an quantized LPC coefficient,
wherein the quantizing includes:
calculating an energy variation of the synthesized voice signal in the previous frames of the input signal;
calculating a changing degree of a sign of the synthesized voice signal in the previous frames of the input signal;
calculating a pitch delay of the synthesized voice signal in the previous frames of the input signal; and
checking whether the synthesized voice signal in the previous frames of the input signal has a voice signal based on the energy variation to perform the first quantization process or the second LSF quantization process, wherein the first quantization process or the second LSF quantization process is performed based on whether the synthesized voice signal has the voice signal, a changing degree of the sign of the synthesized voice signal, and a pitch delay of the synthesized voice signal.
19. A method of selecting dequantization in a voice decoder, comprising:
receiving line spectral frequency (LSF) quantization information of a current frame of an input signal and voice signal synthesis information of the current frame through a specified channel;
generating a synthesized voice signal of a previous frame of the input signal from the voice signal synthesis information of the current frame and LSF quantization information of the previous frame;
selecting, before dequantizing an LSF of the of the current frame, one of a first LSF dequantization process and a second LSF dequantization process for the dequantizing of the LSF of the current frame, wherein the selecting is based on an analysis of the synthesized voice signal;
dequantizing the LSF of the current frame through the selected one of the first dequantization process using a first predictor and the second LSF dequantization process using a second predictor, the second predictor being different from the first predictor, to generate a dequantized LSF vector of the current frame; and
converting the dequantized LSF vector into a dequantized LPC coefficient of the current frame.
20. The method according to claim 19, wherein the dequantizing includes:
calculating an energy variation of the synthesized voice signal of at least the previous frame;
calculating a changing degree of a sign of the synthesized voice signal of at least the previous frame;
calculating a pitch delay of the synthesized voice signal of at least the previous frame; and
checking whether the synthesized voice signal in at least the previous frame has a voice signal based on the calculated energy variation, wherein the one of the first dequatization process and the second dequantization process is selected based on a result of the checking indicating that the synthesized voice signal of at least the previous frame has the voice signal, the calculated changing degree of the sign of the synthesized voice signal of at least the previous frame, and the calculated pitch delay of the synthesized voice signal of at least the previous frame.
21. An apparatus for selecting quantization for a current frame of an input signal in a voice encoder, the apparatus comprising:
an energy calculation unit to calculate respective energy values of subframes of at least a previous frame based upon a synthesized voice signal of at least the previous frame;
an energy buffer to store the calculated energy values;
a moving average calculation unit to calculate two energy moving values based on the stored calculated energy values;
an energy increase calculation unit to calculate an energy increase based on the calculated energy values and the calculated two energy moving values;
an energy decrease calculation unit to calculate an energy decrease based on the calculated energy values and the calculated two energy moving values;
an zero crossing calculation unit to calculate a changing zero crossing rate of the synthesized voice signal;
a pitch difference calculation unit to calculate a difference in a detected pitch delay of the synthesized voice signal; and
a selection signal generation unit to select, before performing quantization of the current frame using any of plural quantization units, which one of the plural quantization units is appropriate for the voice encoding of the current frame based on the synthesized voice signal of at least the previous frame, including consideration of the calculated energy increase, the calculated energy decrease, the calculated zero crossing rate, and the calculated pitch difference.
22. The quantization selection unit according to claim 21, wherein the energy calculation unit calculates respective energy values Ei of ith subframes according to the following equation:
E
i
=
\u2211
n
=
0
L
N
–
1
\u2062
\u2062
s
^
\u2062
\u2062
(
iL
N
+
n
)
2
\u2062
\u2062
i
=
0
,
\u2026
\u2062
,
N
–
1
wherein N is a number of subframes, and L is a number of samples per frame.
23. The quantization selection unit according to claim 21, wherein the energy buffer stores the calculated energy values in a frame unit according to the following equation:
for i=LB\u22121 to 1
EB(i)=EB(i\u22121)
EB(O)=Ei
wherein LB is a length of an energy buffer, and EB is an energy buffer.
24. The quantization selection circuit according to claim 22, wherein the moving average calculation unit calculates two energy moving averages EM,1 and EM,2 according to the following equations:
E
M
,
1
=
1
10
\u2062
\u2211
i
=
5
9
\u2062
\u2062
E
B
\u2061
(
i
)
\u2062
;
and
\u2062
\u2062
E
M
,
2
=
1
10
\u2062
\u2211
i
=
0
9
\u2062
\u2062
E
B
\u2061
(
i
)
.
25. An apparatus for selecting dequantization for a current frame of an input signal in a voice decoder, the apparatus comprising:
an energy calculation unit to calculate respective energy values of subframes of a previous frame of the input signal based on a synthesized voice signal of at least the previous frame;
an energy buffer to store the calculated energy values;
a moving average calculation unit to calculate two energy moving values based on the stored calculated energy values;
an energy increase calculation unit to calculate an energy increase based on the calculated energy values and the calculated two energy moving values;
an energy decrease calculation unit to calculate an energy decrease based on the calculated energy values and the calculated two energy moving values;
an zero crossing calculation unit to calculate a changing zero crossing rate of the synthesized voice signal;
a pitch difference calculation unit to calculate a difference in a detected pitch delay of the synthesized voice signal; and
a selection signal generation unit to generate, before performing dequantization of the current frame using any of plural dequantization units, a selection signal representing a selection of which one of the plural dequantization units is appropriate for the voice encoding of the current frame based on the synthesized voice signal of at least the previous frame, including consideration of the calculated energy increase, the calculated energy decrease, the calculated changing zero crossing rate, and the calculated pitch difference.
26. The dequantization selection unit according to claim 25, wherein the energy calculation unit calculates respective energy values Ei of ith subframes according to the following equation:
E
i
=
\u2211
n
=
0
L
N
–
1
\u2062
\u2062
s
^
\u2062
\u2062
(
iL
N
+
n
)
2
\u2062
\u2062
i
=
0
,
\u2026
\u2062
,
N
–
1
wherein N is a number of subframes, and L is a number of samples per frame.
27. The dequantization selection unit according to claim 25, wherein the energy buffer stores the calculated energy values in a frame unit according to the following equation:
for i=LB\u22121 to 1
EB(i)=EB(i\u22121)
EB(O)=Ei
wherein LB is a length of an energy buffer, and EB is an energy buffer.
28. The dequantization selection circuit according to claim 25, wherein the moving average calculation unit calculates two energy moving averages EM,1 and EM,2 according to the following equations:
E
M
,
1
=
1
10
\u2062
\u2211
i
=
5
9
\u2062
\u2062
E
B
\u2061
(
i
)
\u2062
;
and
\u2062
\u2062
E
M
,
2
=
1
10
\u2062
\u2211
i
=
0
9
\u2062
\u2062
E
B
\u2061
(
i
)
.
29. A voice encoder comprising:
a quantization selection unit checking whether a synthesized voice signal of previous frames of an input signal has a voice signal based on energy variations of the synthesized voice signal of the previous frames of the input signal, and selecting, before quantizing a line spectral frequency (LSF) of a current frame of the input signal, one of a first LSF quantization unit and a second LSF quantization unit for the quantizing of the LSF of the current frame based on a result of the checking indicating that the synthesized voice signal of the previous frames has the voice signal, a changing degree of a sign of the synthesized voice signal, and a pitch delay of the synthesized voice signal of the previous frames; and
a quantization unit quantizing the LSF of the current frame with the selected one of a first LSF quantization unit using a first predictor and the second LSF quantization unit using a second predictor, the second predictor being different from the first predictor, and converting the quantized LSF into a quantized LPC coefficient.
30. A voice encoder comprising:
a quantization selection unit generating a quantization selection signal; and
a quantization unit extracting a linear prediction coding (LPC) coefficient from an input signal, converting the extracted LPC coefficient into a line spectral frequency (LSF), selectively quantizing the LSF with one of a first LSF quantization unit using a first predictor and a second LSF quantization unit using a second predictor, the second predictor being different from the first predictor, based on the quantization selection signal, and converting the quantized LSF into a quantized LPC coefficient,
wherein the quantization selection signal determines the selecting of the one of the first LSF quantization unit and the second LSF quantization unit based on characteristics of a synthesized voice signal in previous frames of the input signal, wherein the LSF is input only to the selected one quantization unit in which the LSF is selectively quantized.