CN109182462B - Method and device for judging whether detection indexes are positive or negative - Google Patents
Method and device for judging whether detection indexes are positive or negative Download PDFInfo
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- CN109182462B CN109182462B CN201811105601.3A CN201811105601A CN109182462B CN 109182462 B CN109182462 B CN 109182462B CN 201811105601 A CN201811105601 A CN 201811105601A CN 109182462 B CN109182462 B CN 109182462B
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- C12Q1/6844—Nucleic acid amplification reactions
Abstract
The invention discloses a method for judging whether a detection index is negative or positive, which comprises the following steps: calculating to obtain a positive control baseline according to the acquired fluorescence data; calculating the positive control base line to obtain a detection index amplification multiple threshold value, and determining an initial value of negative and positive judgment of the detection index according to the amplification multiple threshold value; if the initial value is judged to be negative and the experiment is not finished, determining the state of the detection index to be unknown; if the initial value is judged to be positive, whether the amplification of the experiment is finished is judged, and if the amplification is not finished and the positive discrimination point is smaller than the upper limit value of the preset threshold value, the detection index is judged to be positive; if the amplification is finished, calculating the time value of the inflection point of the fluorescence curve amplification; and judging the state of the obtained detection index according to the time value. According to the invention, the negative and positive detection indexes can be judged without the need of finishing the experiment, and the experiment result can be obtained in advance, so that the negative and positive detection indexes can be judged, and the working efficiency is improved.
Description
Technical Field
The invention relates to the technical field of fluorescence detection, in particular to a method and a device for judging whether a detection index is negative or positive.
Background
Nucleic acid amplification is one of the most important technologies in life science, and isothermal amplification detection is characterized in that the whole process of amplification Reaction is carried out at a single temperature without a special amplifier, and unlike PCR (Polymerase Chain Reaction), the whole process needs to undergo dozens of temperature changes. The characteristics of isothermal amplification technology make their requirements for required instruments simplified and reaction time shortened, because of the great application value.
In the constant-temperature amplification experiment based on fluorescence, the positive and negative judgment of detection indexes is the basis for qualitative and quantitative analysis of nucleic acid amplification technology. Currently, there are various methods for determining the negative or positive of the detection index, for example, calculating the ratio of the fluorescence signal intensity generated by the amplification product of the nucleic acid sample to the initial fluorescence signal intensity of the amplification reaction of the nucleic acid sample, comparing the ratio with a preset threshold value of the ratio of fluorescence signal intensities, and determining the negative or positive of the fluorescence amplification curve according to the comparison result. However, these methods require that the detection index be judged to be negative or positive after all fluorescence data is acquired after the experiment is completed. If the project of simultaneous detection of multiple indexes is detected, the end time of the experiment depends on the index which consumes the longest time, so that the detection time is wasted, and the working efficiency is reduced.
Disclosure of Invention
In order to solve the problems, the invention provides a method and a device for judging whether a detection index is positive or negative, which can judge whether the detection index is positive or negative in real time and improve the working efficiency.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for determining whether a detection index is positive or negative, the method comprising:
calculating to obtain a positive control baseline according to the acquired fluorescence data;
calculating the positive control base line to obtain a detection index amplification multiple threshold value, and determining an initial value of negative and positive judgment of the detection index according to the amplification multiple threshold value;
if the initial value is judged to be negative and the experiment is not finished, determining the state of the detection index to be unknown;
if the initial value is judged to be positive, judging whether the amplification of the experiment is finished, and if the amplification is not finished and the positive judgment point is smaller than the upper limit value of a preset threshold, judging that the detection index is positive;
if the amplification is finished, calculating the time value of the inflection point of the fluorescence curve amplification;
and judging the state of the obtained detection index according to the time value, wherein the state of the detection index comprises unknown, predicted positive, negative, suspected positive and positive.
Optionally, the calculating a positive control baseline from the fluorescence data obtained from the collecting comprises:
acquiring fluorescence data;
generating a fluorescence amplification curve according to the fluorescence data;
and when the fluorescence amplification curve enters an amplification stage, calculating to obtain a positive control baseline according to the current data of the positive control.
Optionally, the calculating the positive control baseline to obtain a detection index amplification multiple threshold, and determining an initial value of negative-positive determination of the detection index according to the amplification multiple threshold includes:
setting an initial amplification multiple threshold value;
correcting the initial amplification threshold according to the form of the positive control baseline to obtain the detection index amplification multiple threshold;
and judging the negative and positive of the detection index according to the amplification multiple threshold value of the detection index to obtain an initial value, wherein the initial value represents the initial negative and positive state of the detection index.
Optionally, the determining whether the amplification is complete comprises:
acquiring fluorescence data of an experiment, and calculating according to the fluorescence data to obtain a first derivative curve;
calculating to obtain the maximum value of the first derivative curve, and obtaining a time point corresponding to the maximum value;
and if points meeting the preset fluorescence value exist on two sides of the time point, judging that the experimental amplification is finished.
Optionally, the determining, according to the time value, a state of obtaining the detection index includes:
judging whether the time value is larger than an upper limit value of a preset threshold value or not, and if so, determining the detection index as negative;
if not, judging whether the time value is smaller than a lower limit value of a preset threshold value, if so, determining the detection index to be positive, and if not, determining the detection index to be suspected positive.
A determination device for detecting whether an index is positive or negative, comprising:
the first calculation unit is used for calculating and obtaining a positive control baseline according to the acquired fluorescence data;
the second calculation unit is used for calculating the positive control base line to obtain a detection index amplification multiple threshold value, and determining an initial value of negative and positive judgment of the detection index according to the amplification multiple threshold value; a first judging unit, configured to determine that the state of the detection index is unknown if the initial value is determined to be negative and the experiment is not completed;
a second judging unit, configured to judge whether the amplification of the experiment is completed if the initial value is judged to be positive, and judge that the detection indicator is predictive positive if the amplification is not completed and a positive judgment point is smaller than an upper limit of a preset threshold;
the third calculation unit is used for calculating the time value of the inflection point of the fluorescence curve amplification if the amplification is finished;
and the third judging unit is used for judging the state of the obtained detection index according to the time value, wherein the state of the detection index comprises unknown, predicted positive, negative, suspected positive and positive.
Optionally, the first computing unit includes:
the acquisition subunit is used for acquiring fluorescence data;
a generating subunit, configured to generate a fluorescence amplification curve according to the fluorescence data;
and the first calculating subunit is used for calculating to obtain a positive control baseline according to the current data of the positive control when the fluorescence amplification curve enters the amplification stage.
Optionally, the second computing unit includes:
a setting subunit, configured to set an initial amplification factor threshold;
a correction subunit, configured to correct the initial amplification threshold according to a form of the positive control baseline, so as to obtain the detection index amplification multiple threshold;
and the judging subunit is used for judging the negative and positive of the detection index according to the detection index amplification multiple threshold value to obtain an initial value, wherein the initial value represents the initial negative and positive state of the detection index.
Optionally, the apparatus further comprises an amplification determination unit, the amplification determination unit comprising:
the curve calculation subunit is used for acquiring fluorescence data of an experiment and calculating to obtain a first derivative curve according to the fluorescence data;
the second calculating subunit is used for calculating and obtaining the maximum value of the first-order derivative curve and obtaining a time point corresponding to the maximum value;
and the amplification judging subunit is used for judging that the experimental amplification is finished if points meeting a preset fluorescence value exist on two sides of the time point.
Optionally, the third judging unit includes:
the first judgment subunit is used for judging whether the time value is larger than an upper limit value of a preset threshold value or not, and if so, determining the detection index as negative;
and the second judging subunit is used for judging whether the time value is smaller than a lower limit value of a preset threshold value or not if the time value is not smaller than the lower limit value of the preset threshold value, determining the detection index to be positive if the time value is smaller than the lower limit value of the preset threshold value, and determining the detection index to be suspected positive if the time value is not smaller than the lower limit value of the preset threshold value.
Compared with the prior art, the invention provides the method and the device for judging the negative and positive detection indexes, the fluorescence data is acquired in real time, the positive control base line is obtained by calculation, the initial value of the detection indexes can be determined by determining the amplification multiple threshold value according to the control base line, and then the negative and positive detection indexes can be judged in real time according to the amplification multiple threshold value in the amplification stage, so that the negative and positive detection indexes can be judged without judging the negative and positive detection indexes after the experiment is finished, the experiment result can be obtained in advance, the negative and positive detection indexes can be judged, and the working efficiency is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a schematic flow chart of a method for determining whether a detected indicator is positive or negative according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a fluorescence amplification curve provided in an embodiment of the present invention;
FIG. 3 is a schematic flow chart of a method for determining whether amplification is complete according to an embodiment of the present invention;
fig. 4 is a schematic structural diagram of a determination device for detecting whether an index is positive or negative according to an embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The terms "first" and "second," and the like in the description and claims of the present invention and the above-described drawings are used for distinguishing between different objects and not for describing a particular order. Furthermore, the terms "comprising" and "having," as well as any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, system, article, or apparatus that comprises a list of steps or elements is not set forth for a listed step or element but may include steps or elements not listed.
In the embodiment of the present invention, a method for determining whether a detection indicator is positive or negative is provided, referring to fig. 1, including:
and S101, calculating to obtain a positive control baseline according to the acquired fluorescence data.
The fluorescence data is collected in real time to generate a fluorescence amplification curve, when the fluorescence curve enters an amplification period, a positive control baseline is calculated according to the current data of the positive control, and referring to fig. 2, the schematic diagram of the fluorescence amplification curve provided by the embodiment of the invention is shown.
S102, calculating the positive control base line to obtain a detection index amplification multiple threshold value, and determining an initial value of negative and positive judgment of the detection index according to the amplification multiple threshold value.
The process of calculating the detection index amplification multiple threshold value comprises the following steps:
setting an initial amplification multiple threshold value;
and correcting the initial amplification threshold according to the form of the positive control baseline to obtain the detection index amplification multiple threshold.
Specifically, when the detection index is a positive control index, the set fold amplification threshold value TV1 and the set fold amplification threshold value TV2 are used.
And when the detection index is a non-positive control index, acquiring a baseline of the positive control curve. When the baseline is less than the set threshold, the set fold amplification threshold TV1 and the fold amplification threshold TV2 are used. When the baseline is greater than or equal to the set threshold, the positive and negative of the positive control curve are judged (the judging process is consistent with the main flow judging positive and negative process).
If the positive control curve is negative, the set fold amplification threshold value TV1 and the fold amplification threshold value TV2 are used.
And if the positive control curve is positive, calculating the amplification multiple ER of the positive control curve. Correcting the fold amplification threshold value TV1 to the fold amplification ER of the positive control curve if the fold amplification ER of the positive control curve is greater than the set threshold value and less than the set threshold value TV 1; if the fold amplification ER of the positive control curve is greater than the set threshold and less than the set threshold TV2, the fold amplification threshold TV2 is corrected to the fold amplification ER of the positive control curve. Otherwise, the set amplification factor threshold value TV1 and the amplification factor threshold value TV2 are used.
The initial value of the positive-negative determination of the detection index, namely the current positive-negative of the detection index, can then be determined according to the amplification factor threshold.
S103, if the initial value is judged to be negative and the experiment is not finished, determining the state of the detection index to be unknown;
s104, if the initial value is judged to be positive, judging whether the amplification of the experiment is finished or not, and if the amplification is not finished and the positive judgment point is smaller than the upper limit value of a preset threshold, judging that the detection index is positive;
s105, if the amplification is finished, calculating the time value of the inflection point of the fluorescence curve amplification;
and S106, judging and obtaining the state of the detection index according to the time value, wherein the state of the detection index comprises unknown, predicted positive, negative, suspected positive and positive.
It should be noted that the detection index states are divided into five states: unknown, negative, positive, predictive positive and suspected positive. Wherein, the unknown represents the initial state, and the positive and negative of the detection index can not be distinguished at present; negative representation determines that the detection index is negative, positive representation determines that the detection index is positive, prediction positive representation indicates that the detection index is likely positive, and multiple data support is needed; a suspected positive is an indication that the test cannot be determined to be positive, and further determination is required.
When the detection index state is judged in real time, if the index is judged to be negative and the experiment is not finished at the moment, the index state is still displayed as unknown; if the index is positive, judging whether the amplification is finished.
The embodiment of the present invention further provides a method for determining whether amplification is completed, and with reference to fig. 3, the method includes:
s301, acquiring fluorescence data of an experiment, and calculating according to the fluorescence data to obtain a first derivative curve;
s302, calculating to obtain the maximum value of the first derivative curve, and obtaining a time point corresponding to the maximum value;
s303, if points meeting a preset fluorescence value exist on two sides of the time point, judging that the experimental amplification is finished.
Specifically, the first derivative curve is calculated from the current fluorescence data, and the maximum value of the first derivative curve is calculated. The first derivative is calculated as:
wherein x isiRepresenting the time value of the i-th cycle, yiIndicating the fluorescence value for the ith cycle, and N identifies the current cycle number.
And (3) taking the maximum value m of the first derivative and the corresponding time point t, and respectively searching points with fluorescence values meeting the preset fluorescence value range on two sides of t, wherein the preset fluorescence value is 0.1 × m, if the points exist on two sides, the amplification is finished, otherwise, the amplification is not finished.
If the amplification is finished, calculating the time value T of the inflection point of the fluorescence amplification curvep。
According to TpThe value judgment index state is as follows:
the method comprises the steps that S marks the current state of a detection index, Cutpff represents a preset threshold value, UpCutpff represents an upper limit value of the preset threshold value, and LowCutpff represents a lower limit value of the preset threshold value, wherein the preset threshold value is determined based on analysis summary of historical data.
The invention provides a method for judging whether a detection index is negative or positive, which comprises the steps of acquiring fluorescence data in real time, calculating to obtain a positive control baseline, determining an amplification multiple threshold value according to the control baseline to determine an initial value of the detection index, and then judging whether the detection index is negative or positive in real time according to the amplification multiple threshold value in an amplification stage, so that the detection index is negative or positive without judging whether the detection index is negative or positive after the experiment is finished, an experiment result can be obtained in advance, the detection index is negative or positive, and the working efficiency is improved.
The embodiment of the invention also provides a method for calculating the time value of the fluorescence curve amplification inflection point, wherein the fluorescence amplification curves with different morphological characteristics are classified for calculating the fluorescence curve amplification inflection point, and then different inflection point calculation methods can be adopted for different classification curves.
Specifically, whether the curve is a standard negative curve is judged according to the maximum value of a first derivative distribution curve of the fluorescence amplification curve and the amplification multiple of the fluorescence curve, and if the maximum value is lower than a set negative threshold value and the amplification multiple is smaller than a set amplification multiple threshold value, the curve is judged to be a negative curve. The negative curve has no inflection point. The fold amplification calculation formula is as follows:
wherein the content of the first and second substances,
the N point average value taking the maximum value of the curve as a core is obtained, and the value range of N is 3-7.
The average value of the N points with the minimum value of the curve as the core is shown.
And if the maximum value of the first derivative distribution curve is between the negative threshold and the positive threshold, calculating the amplitude of the first derivative distribution curve, and if the amplitude is greater than the set amplitude threshold, judging that the first derivative distribution curve is not the S characteristic curve. The non-S characteristic curve is a negative curve and has no inflection point. The amplitude calculation formula is as follows:
β＝X2(0.1×ymax)-X1(0.1×ymax)
wherein, X1(0.1 Xy)max) The time corresponding to the maximum value 0.1 times to the left of the maximum value of the curve.
X2(0.1×ymax) The time corresponding to the maximum value 0.1 times to the right of the maximum value of the curve.
And if the maximum value of the first derivative distribution curve is larger than the positive threshold value, searching whether a secondary maximum value exists before the maximum value of the second derivative distribution curve, and if the secondary maximum value exists and the corresponding first derivative distribution curve in the range of the secondary maximum value and the maximum value is monotonically increased, judging that the second type S curve is the second type S curve. Such a curve takes the moment corresponding to the second maximum of the second derivative distribution curve as the inflection point.
And if the maximum value of the first derivative is larger than the positive threshold value and the S-curve two-type characteristic does not exist, calculating the skewness of the curve, and if the skewness is larger than the set skewness threshold value, judging the S-curve three-type characteristic. The skewness calculation formula is as follows:
α＝|2X(ymax)-X1(0.1×ymax)-X2(0.1×ymax)|
wherein, X (y)max) The moment corresponding to the maximum value of the curve.
The curve firstly determines a preliminary amplification period range according to first derivative distribution, a minimum maximum fluorescence signal value is searched in the amplification range, and the time corresponding to the minimum fluorescence value is accumulated by a certain multiple of the difference value of the minimum maximum fluorescence value and serves as an inflection point.
If the above characteristics do not exist, the S-curve I type is judged. Such a curve takes the time corresponding to the maximum of the second derivative distribution curve as an inflection point.
The invention provides a method for judging and displaying the detection index state in real time, which can synchronously update the currently judged detection index state in an experiment. The user can obtain the relevant information at the first time, so that the waiting time of the user is reduced, and the efficiency is improved. Particularly for the index with high amplification speed, the positive index determination time is greatly advanced. For example, the time length of the whole experimental amplification stage is 50 minutes, and the method can judge the positive index by more than ten minutes or even more than twenty minutes in advance for certain indexes, thereby shortening the time by nearly one time, and having great practical value in application fields with higher time requirements, such as entry-exit inspection and the like.
The embodiment of the present invention further provides a device for determining whether an index is positive or negative, referring to fig. 4, including:
a first calculating unit 401, configured to calculate a positive control baseline according to the acquired fluorescence data;
a second calculating unit 402, configured to calculate the positive control baseline, obtain a detection index amplification multiple threshold, and determine an initial value of negative-positive determination of the detection index according to the amplification multiple threshold;
a first determining unit 403, configured to determine that the state of the detection index is unknown if the initial value is determined to be negative and the experiment is not completed;
a second determination unit 404, configured to determine whether the amplification of the experiment is completed if the initial value is determined to be positive, and determine that the detection indicator is predictive positive if the amplification is not completed and a positive determination point is smaller than an upper limit of a preset threshold;
a third calculating unit 405, configured to calculate a time value of an inflection point of the fluorescence curve amplification if the amplification is completed;
a third determining unit 406, configured to determine, according to the time value, a state of obtaining the detection index, where the state of the detection index includes unknown, predicted positive, negative, suspected positive, and positive.
Optionally, the first computing unit includes:
the acquisition subunit is used for acquiring fluorescence data;
a generating subunit, configured to generate a fluorescence amplification curve according to the fluorescence data;
and the first calculating subunit is used for calculating to obtain a positive control baseline according to the current data of the positive control when the fluorescence amplification curve enters the amplification stage.
Optionally, the second computing unit includes:
a setting subunit, configured to set an initial amplification factor threshold;
a correction subunit, configured to correct the initial amplification threshold according to a form of the positive control baseline, so as to obtain the detection index amplification multiple threshold;
and the judging subunit is used for judging the negative and positive of the detection index according to the detection index amplification multiple threshold value to obtain an initial value, wherein the initial value represents the initial negative and positive state of the detection index.
Optionally, the apparatus further comprises an amplification determination unit, the amplification determination unit comprising:
the curve calculation subunit is used for acquiring fluorescence data of an experiment and calculating to obtain a first derivative curve according to the fluorescence data;
the second calculating subunit is used for calculating and obtaining the maximum value of the first-order derivative curve and obtaining a time point corresponding to the maximum value;
and the amplification judging subunit is used for judging that the experimental amplification is finished if points meeting a preset fluorescence value exist on two sides of the time point.
Optionally, the third judging unit includes:
the first judgment subunit is used for judging whether the time value is larger than an upper limit value of a preset threshold value or not, and if so, determining the detection index as negative;
and the second judging subunit is used for judging whether the time value is smaller than a lower limit value of a preset threshold value or not if the time value is not smaller than the lower limit value of the preset threshold value, determining the detection index to be positive if the time value is smaller than the lower limit value of the preset threshold value, and determining the detection index to be suspected positive if the time value is not smaller than the lower limit value of the preset threshold value.
The invention provides a judgment device for detecting the negative and positive indexes, which is characterized in that fluorescence data is acquired in real time in a first calculation unit, a positive control baseline is calculated and acquired, and then the negative and positive indexes of the detection indexes can be judged in real time according to an amplification multiple threshold value in an amplification stage through a second calculation unit, a first judgment unit, a second judgment unit, a third calculation unit and a third judgment unit, so that the negative and positive indexes of the detection indexes can be judged in advance without judging the negative and positive indexes after the experiment is finished, the experiment result can be acquired in advance, the negative and positive indexes of the detection indexes can be judged, and the working efficiency is improved.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. A method for determining whether a detection index is positive or negative, the method comprising:
calculating to obtain a positive control baseline according to the acquired fluorescence data;
correcting and calculating the set initial amplification multiple threshold value by using the positive control baseline to obtain a detection index amplification multiple threshold value, and determining the initial value of negative and positive judgment of the detection index according to the amplification multiple threshold value;
if the initial value is judged to be negative and the experiment is not finished, determining the state of the detection index to be unknown;
if the initial value is judged to be positive, judging whether the amplification of the experiment is finished, and if the amplification is not finished and the positive judgment point is smaller than the upper limit value of a preset threshold, judging that the detection index is positive;
if the amplification is finished, calculating the time value of the inflection point of the fluorescence curve amplification according to the type of the fluorescence curve;
judging the state of the detection index according to the time value, wherein the state of the detection index comprises unknown, predicted positive, negative, suspected positive and positive;
the judging whether the amplification of the experiment is finished comprises the following steps:
acquiring fluorescence data of an experiment, and calculating according to the fluorescence data to obtain a first derivative curve;
calculating to obtain the maximum value of the first derivative curve, and obtaining a time point corresponding to the maximum value;
if points meeting a preset fluorescence value exist on two sides of the time point, judging that the experimental amplification is finished;
the determining the state of obtaining the detection index according to the time value includes:
judging whether the time value is larger than an upper limit value of a preset threshold value or not, and if so, determining the detection index as negative;
if not, judging whether the time value is smaller than a lower limit value of a preset threshold value, if so, determining the detection index to be positive, and if not, determining the detection index to be suspected positive.
2. The method of claim 1, wherein calculating a positive control baseline from the fluorescence data obtained from the acquisition comprises:
acquiring fluorescence data;
generating a fluorescence amplification curve according to the fluorescence data;
and when the fluorescence amplification curve enters an amplification stage, calculating to obtain a positive control baseline according to the current data of the positive control.
3. The method of claim 1, wherein the performing a correction calculation on the set initial amplification factor threshold using the positive control baseline to obtain a detection index amplification factor threshold, and determining an initial value of a negative-positive determination of the detection index according to the amplification factor threshold comprises:
setting an initial amplification multiple threshold value;
correcting the initial amplification threshold according to the form of the positive control baseline to obtain the detection index amplification multiple threshold;
and judging the negative and positive of the detection index according to the amplification multiple threshold value of the detection index to obtain an initial value, wherein the initial value represents the initial negative and positive state of the detection index.
4. A determination device for detecting whether an index is positive or negative, comprising:
the first calculation unit is used for calculating and obtaining a positive control baseline according to the acquired fluorescence data;
the second calculation unit is used for performing correction calculation on the set initial amplification multiple threshold value by using the positive control baseline to obtain a detection index amplification multiple threshold value, and determining an initial value of negative and positive judgment of the detection index according to the amplification multiple threshold value;
a first judging unit, configured to determine that the state of the detection index is unknown if the initial value is determined to be negative and the experiment is not completed;
a second judging unit, configured to judge whether the amplification of the experiment is completed if the initial value is judged to be positive, and judge that the detection indicator is predictive positive if the amplification is not completed and a positive judgment point is smaller than an upper limit of a preset threshold;
the third calculation unit is used for calculating the time value of the inflection point of the amplification of the fluorescence curve according to the type of the fluorescence curve if the amplification is finished;
a third judging unit, configured to judge, according to the time value, a state of the detection index, where the state of the detection index includes unknown, predicted positive, negative, suspected positive, and positive;
the apparatus further includes an amplification judging unit including:
the curve calculation subunit is used for acquiring fluorescence data of an experiment and calculating to obtain a first derivative curve according to the fluorescence data;
the second calculating subunit is used for calculating and obtaining the maximum value of the first-order derivative curve and obtaining a time point corresponding to the maximum value;
an amplification judging subunit, configured to judge that the experimental amplification is completed if there are points satisfying a preset fluorescence value on both sides of the time point;
the third judging unit includes:
the first judgment subunit is used for judging whether the time value is larger than an upper limit value of a preset threshold value or not, and if so, determining the detection index as negative;
and the second judging subunit is used for judging whether the time value is smaller than a lower limit value of a preset threshold value or not if the time value is not smaller than the lower limit value of the preset threshold value, determining the detection index to be positive if the time value is smaller than the lower limit value of the preset threshold value, and determining the detection index to be suspected positive if the time value is not smaller than the lower limit value of the preset threshold value.
5. The apparatus of claim 4, wherein the first computing unit comprises:
the acquisition subunit is used for acquiring fluorescence data;
a generating subunit, configured to generate a fluorescence amplification curve according to the fluorescence data;
and the first calculating subunit is used for calculating to obtain a positive control baseline according to the current data of the positive control when the fluorescence amplification curve enters the amplification stage.
6. The apparatus of claim 4, wherein the second computing unit comprises:
a setting subunit, configured to set an initial amplification factor threshold;
a correction subunit, configured to correct the initial amplification threshold according to a form of the positive control baseline, so as to obtain the detection index amplification multiple threshold;
and the judging subunit is used for judging the negative and positive of the detection index according to the detection index amplification multiple threshold value to obtain an initial value, wherein the initial value represents the initial negative and positive state of the detection index.
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| CN111161338B (en) * | 2019-12-26 | 2022-05-17 | 浙江大学 | Point cloud density improving method for depth prediction based on two-dimensional image gray scale |
| CN112763470A (en) * | 2020-12-28 | 2021-05-07 | 季华实验室 | Multi-channel bioluminescence detection method |
| CN112730369A (en) * | 2021-01-25 | 2021-04-30 | 苏州市中心血站 | Multichannel fluorescence detection data processing method |
| CN114444927A (en) * | 2021-02-02 | 2022-05-06 | 方舟生物安全科技(广州)有限公司 | Pathogenic microorganism Internet of things real-time monitoring system based on high risk area division |
| CN114058683B (en) * | 2021-11-17 | 2022-09-30 | 杭州优思达生物技术有限公司 | Method for judging nucleic acid amplification result |
| CN114480744B (en) * | 2022-03-11 | 2022-11-11 | 深圳市研元生物科技有限公司 | New coronavirus isothermal detection kit based on CRISPR-Cas system, use method and application |
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| US7991558B2 (en) * | 2005-09-29 | 2011-08-02 | Roche Molecular Systems, Inc. | Systems and methods for determining real-time PCR cycle thresholds using cluster analysis |
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