CN104561322A - Quantitative gastric carcinoma patient survival prediction and individual follow-up schedule evaluation method based on helicobacter pylori DNA molecules - Google Patents
Quantitative gastric carcinoma patient survival prediction and individual follow-up schedule evaluation method based on helicobacter pylori DNA molecules Download PDFInfo
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Abstract
The invention discloses a quantitative gastric carcinoma patient survival prediction and individual follow-up schedule evaluation method based on helicobacter pylori DNA molecules. The survival prediction method comprises the following steps: (1) detecting the relative infection dose of helicobacter pylori (Hp) in gastric carcinoma tissues; and (2) performing prognosis evaluation according to the detected relative infection dose of helicobacter pylori, wherein the poor prognosis degree is increased along with increase of the relative infection dose of helicobacter pylori. According to the method, the prognosis of a gastric carcinoma patient is evaluated so as to determine the most reasonable follow-up schedule suitable for the patient, and according to follow-up at the reasonable follow-up schedule, metastasis or reoccurrence symptoms are early discovered, and the survival quality and survival schedule of the gastric carcinoma patients are improved.
Description
Technical field
The present invention relates to patients with gastric cancer post-surgical condition electric powder prediction, particularly relate to a kind of patients with gastric cancer Prediction of survival quantitative based on Hp DNA molecular and individuation follow up time appraisal procedure.
Background technology
According to the recent statistics data presentation of world's cancer research tissue (IARC) Globocan, within 2012, global cancer of the stomach neopathy number is 95.2 more than ten thousand people, accounts for 6.8% of global pathogenesis of cancer number, occupy the 5th; The death toll of whole world cancer of the stomach is 72.3 more than ten thousand people, accounts for 8.8% of the total lethal number of global cancer, is positioned at the 3rd.Cancer is a kind of disease of specific type, and cancer patients accepts after effective regular radical treatment in hospital, clinical recovery rehabilitation and leaving hospital, but cancer patients deposits the possibility of recurrence or transfer all the time, needs long term follow-up to cancer patient.Therefore reasonably the formulation of follow up time seems particularly important: one, follow up a case by regular visits to too short meeting interval time and cause unnecessary rechecking to be lost time and financial resources; Two, cause can not the tumour sign that shifts in early days of Timeliness coverage for the too short meeting in follow up time interval, causes the state of an illness to be incured loss through delay, is unfavorable for the prognosis of patient.In US National comprehensive cancer network (National Comprehensive Cancer Network, NCCN) guide 2013 the 2nd edition, the follow up time of patients with gastric cancer is mentioned like this: all patients with gastric cancer all should accept following up a case by regular visits to of system.Follow up a case by regular visits to content and comprise comprehensive disease history inquire and physical examination, within every 3-6 month, follow up a case by regular visits to 1 time, 1-2 altogether; Within every 6-12 month afterwards, follow up a case by regular visits to 1 time, 3-5 altogether; Annual 1 time later; Carry out CBC, biochemical index, imaging examination or endoscopy according to clinical setting simultaneously.Highlight the importance of following up a case by regular visits in guide, but do not provide how to follow up a case by regular visits to scheme according to the individuation of patient self state of an illness.
Summary of the invention
In view of this, the embodiment of the present invention provides a kind of patients with gastric cancer Prediction of survival quantitative based on Hp DNA molecular and individuation follow up time appraisal procedure, main purpose estimates to formulate the most reasonable follow up time of applicable patient, and following up a case by regular visits to by reasonable follow up time, early discovery transfer or recurrence sign, improve life quality and the survival time of patients with gastric cancer.
For achieving the above object, the present invention mainly provides following technical scheme:
On the one hand, embodiments provide a kind of patients with gastric cancer Prediction of survival method quantitative based on Hp DNA molecular, comprise the steps:
1) the relative infective dose of Hp (Hp) in stomach organization is detected;
2) prognosis evaluation is carried out according to the relative infective dose of the Hp detected;
Wherein prognosis mala degree increases with the rising of the relative infective dose of Hp.
As preferably, the detection of the relative infective dose of Hp adopts real-time fluorescence quantitative PCR (qRT-PCR).
As preferably, reaction conditions is, 95 DEG C of denaturation 5min, 95 DEG C of sex change 10s, and 60 DEG C of annealing extend 30s, 40 circulations.
As preferably, the relative infective dose of Hp is obtained by following formula, relative infective dose=2 of Hp
-△ △ Ct, wherein △ △ Ct=(Ct sample object-Ct sample internal reference)-(Ct blank group object-Ct blank group internal reference), selects blank group to be negative findings for standard with methylene blue dyeing and regular-PCR; The relative infective dose of blank group is 1, namely≤1 is that HP is negative, and > 1 is that Hp is positive, and experiment repetition is averaged for 3 times.
On the other hand, embodiments provide a kind of patients with gastric cancer individuation follow up time appraisal procedure quantitative based on Hp DNA molecular, comprise the steps:
1) the relative infective dose of Hp in stomach organization is detected;
2) term of reference of the relative infective dose of patients with gastric cancer prognosis mala Hp is formulated;
3) the relative infective dose of the Hp of detection and gained term of reference are compared, determine the scheme of following up a case by regular visits to.
As preferably, described term of reference is as follows: Helicobacter pylori infection negative patient, when namely the relative infective dose of Hp is less than or equal to 1, without the need to special follow up time, is undertaken by international guidelines: within every 3-6 month, follow up a case by regular visits to 1 time, 1-2 altogether; Within every 6-12 month afterwards, follow up a case by regular visits to 1 time, 3-5 altogether; Annual 1 time later;
Helicobacter pylori infection positive patient, namely when the relative infective dose of Hp is greater than 1, after chemotherapy, recommend the radical cure treatment carrying out Hp, the experiment and periodic monitoring C14 exhales, and follow up time should carry out corresponding shortening with Hp relative to infective dose:
During 1 < Hp relative infective dose < 15, within postoperative 3 years every 3-4 month, follow up a case by regular visits to once, at least follow up a case by regular visits to 3 times every year; 4-5 follows up a case by regular visits to once every half a year;
Relative infective dose >=15 of Hp, follow up a case by regular visits to once, at least follow up a case by regular visits to 4 times every year in postoperative 3 years every 2-3 month; 4-5 follows up a case by regular visits to once for every 4 months.
As preferably, the radical cure treatment plan of Hp is as follows: adopt standard triple therapy, i.e. PPI+ clarithromycin+amoxycilline Trihydrate bp or PPI+ clarithromycin+metronidazole, the course for the treatment of is 10 days or 14 days.
As preferably, the detection of the relative infective dose of Hp adopts real-time fluorescence quantitative PCR (qRT-PCR).
As preferably, reaction conditions is, 95 DEG C of denaturation 5min, 95 DEG C of sex change 10s, and 60 DEG C of annealing extend 30s, 40 circulations.
As preferably, the relative infective dose of Hp is obtained by following formula, relative infective dose=2 of Hp
-△ △ Ct, wherein △ △ Ct=(Ct sample object-Ct sample internal reference)-(Ct blank group object-Ct blank group internal reference), selects blank group to be negative findings for standard with methylene blue dyeing and regular-PCR; The relative infective dose of blank group is 1, namely≤1 is that HP is negative, and > 1 is that Hp is positive, and experiment repetition is averaged for 3 times.
Compared with prior art, beneficial effect of the present invention is:
1., in the Forecasting Methodology of the embodiment of the present invention, the real time fluorescent quantitative of use detects the relative infective dose technology maturation of Hp, is widely used in clinical detection and laboratory study in countries in the world;
2. in the Forecasting Methodology of the embodiment of the present invention, the patient's (research according to us about has 25%) detecting Hp feminine gender follows up a case by regular visits to without the need to special reinforcement, greatly can reduce patients ' psychological burden;
3. in the Forecasting Methodology of the embodiment of the present invention, according to the follow up time of Hp relative infective dose adjustment patient, avoid follow up time interval because of unitized and can not the tumour sign that shifts in early days of Timeliness coverage, the state of an illness is caused to be incured loss through delay, lose best occasion for the treatment, thus result for the treatment of is bad, causes that sb.'s illness took a turn for the worse, be unfavorable for patient's prognosis, realize the high performance-price ratio for the treatment of and prevention of tumour;
4. in the Forecasting Methodology of the embodiment of the present invention, detect Hp positive patient and patient can be reminded to pay close attention to own bodies change, in order to avoid delay treatment, for early discovery and these diseases for the treatment of are laid a good foundation;
5. in the Forecasting Methodology of the embodiment of the present invention, negative and positive according to Hp, and the number of infective dose, patient is divided into different grades, formulates different follow up time according to different grades, namely Hp infective dose is higher, it is shorter to follow up a case by regular visits to interval time, not only can changing by Timeliness coverage conditions of patients, extend patient's life-span, can also be individual and country's a large amount of human and material resources of saving and financial resources.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet of following up a case by regular visits to embodiment of the embodiment of the present invention.
Fig. 2 A-Fig. 2 D is that different Hp detection method detects the relative infective dose of Hp to the impact of patients with gastric cancer prognosis.
Fig. 3 A is that patients with gastric cancer adds up survival curve, and Fig. 3 B is different times survival and Died Patients Hp infective dose graph of a relation.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail, but not as a limitation of the invention.In the following description, the not necessarily same embodiment that different " embodiment " or " embodiment " refers to.In addition, special characteristic, structure or feature in one or more embodiment can be combined by any suitable form.
With reference to Fig. 1, based on the patients with gastric cancer Prediction of survival method that Hp DNA molecular is quantitative, comprise the steps:
1) the relative infective dose of Hp (Hp) in stomach organization is detected;
2) prognosis evaluation is carried out according to the relative infective dose of the Hp detected;
Wherein prognosis mala degree increases with the rising of the relative infective dose of Hp.
The prognosis of relative infective dose to patients with gastric cancer by detecting Hp (Hp) in stomach organization in the embodiment of the present invention is estimated.More adequately can predict whether are the survival rate of patients with gastric cancer and prognosis mala.And formulate the most reasonable follow up time being applicable to patient on this basis, and following up a case by regular visits to by reasonable follow up time, early discovery transfer or recurrence sign, improve life quality and the survival time of patients with gastric cancer.
Preferred as above-described embodiment, the detection of the relative infective dose of Hp adopts real-time fluorescence quantitative PCR (qRT-PCR).The method surveys the method for each polymerase chain reaction (PCR) circulation after product total amount with fluorescent chemical, strong interference immunity, and prediction accurately.The method technology is comparatively ripe, easy handling.
The reaction system of the method is as follows:
The reaction conditions of the method is, 95 DEG C of denaturation 5min, 95 DEG C of sex change 10s, and 60 DEG C of annealing extend 30s, 40 circulations.The relative infective dose of Hp is obtained by following formula, relative infective dose=2 of Hp
-△ △ Ct, wherein △ △ Ct=(Ct sample object-Ct sample internal reference)-(Ct blank group object-Ct blank group internal reference), selects blank group to be negative findings for standard with methylene blue dyeing and regular-PCR; The relative infective dose of blank group is 1, namely≤1 is that HP is negative, and > 1 is that Hp is positive, and experiment repetition is averaged for 3 times.
On the other hand, with reference to Fig. 1, embodiments provide a kind of patients with gastric cancer individuation follow up time appraisal procedure quantitative based on Hp DNA molecular, comprise the steps:
1) the relative infective dose of Hp in stomach organization is detected;
2) term of reference of the relative infective dose of patients with gastric cancer prognosis mala Hp is formulated;
3) the relative infective dose of the Hp of detection and gained term of reference are compared, determine the scheme of following up a case by regular visits to.
The detection of the relative infective dose of Hp adopts the method for above-described embodiment.
Preferred as above-described embodiment, described term of reference is as follows: Helicobacter pylori infection negative patient, when namely the relative infective dose of Hp is less than or equal to 1, without the need to special follow up time, is undertaken by international guidelines: within every 3-6 month, follow up a case by regular visits to 1 time, 1-2 altogether; Within every 6-12 month afterwards, follow up a case by regular visits to 1 time, 3-5 altogether; Annual 1 time later;
Helicobacter pylori infection positive patient, namely when the relative infective dose of Hp is more than or equal to 1, after chemotherapy, recommend the radical cure treatment carrying out Hp, the experiment and periodic monitoring C14 exhales, and follow up time should carry out corresponding shortening with Hp relative to infective dose:
During 1 < Hp relative infective dose < 15, within postoperative 3 years every 3-4 month, follow up a case by regular visits to once, at least follow up a case by regular visits to 3 times every year; 4-5 follows up a case by regular visits to once every half a year;
Relative infective dose >=15 of Hp, follow up a case by regular visits to once, at least follow up a case by regular visits to 4 times every year in postoperative 3 years every 2-3 month; 4-5 follows up a case by regular visits to once for every 4 months.
Preferred as above-described embodiment, the radical cure treatment plan of Hp is as follows: adopt standard triple therapy, i.e. PPI+ clarithromycin+amoxycilline Trihydrate bp or PPI+ clarithromycin+metronidazole, the course for the treatment of is 10 days or 14 days.
Fig. 2 A-Fig. 2 D is that different Hp detection method detects the relative infective dose of Hp to the impact of patients with gastric cancer prognosis, and wherein Fig. 2 A is methylene blue staining; Fig. 2 B is regular-PCR detection method; Fig. 2 C and Fig. 2 D is real-time fluorescence quantitative PCR detection method; As can be seen from the figure, methylene blue dyeing read tablet unstable and the drawback such as regular-PCR band interfering factors is many, therefore the embodiment of the present invention selects real-time fluorescence quantitative PCR detection method as Hp method of detecting infection.Fig. 2 C display and Hp infect positive patient comparatively negative patient's poor prognosis; It is higher that Fig. 2 D shows Hp infective dose, patient's poor prognosis.
For 118 routine cancer of the stomach Hp positive patient samples:
1. first infect positive patient to the postoperative gastric cancer Hp of clinical certain one-phase (for successive years) to follow up a case by regular visits to (3-5), obtain Follow-up Data.
2., then by the Hp infective dose typing SPSS 17.0 statistical analysis software of follow-up of patients's data and mensuration, make the database of this sample.
3. in this database, filter out the data (59 example) of half patient dead at first, mean (the mean of the Hp infective dose of this 59 routine patient is analyzed in SPSS software, M), standard deviation (standard deviation, SD) and each Hp infect grade (Hp≤1,1 < Hp < 15, Hp >=15) distribution.Adopt the estimation range formulating the Hp infective dose that patients with gastric cancer Postoperative determination is estimated, the scope of the M=9.75 of this 59 routine patient, SD=6.45, gained Hp infective dose is 0.12-26.76.
4. in SPSS software, make patients with gastric cancer (118 example) total survival curve, find out time during residue 50% living patients, be the prediction half patient of Hp infective dose, i.e. 50% death time.
Fig. 3 A is that patients with gastric cancer adds up survival curve, Fig. 3 B is different times survival and Died Patients Hp infective dose graph of a relation, wherein Fig. 3 A is the accumulative existence of 118 routine patients with gastric cancer (CumSurvival) curve, the Hp being left 50% in this curve 24 months time infects positive patient survival, the scope of Hp infective dose is (0.12,-26.76), in half patient dead at first, it is 79.4% that Hp infects positive rate, negative rate is 20.3% (and in remaining half patient, it is 71.2% that Hp infects positive rate, and negative rate is 28.8%).The meaning of this Forecasting Methodology is: if the Hp of patients with gastric cancer infects positive, then this patients with gastric cancer after radical operation had dead possibility to be 79.4% in 24 months.
Shown in Fig. 3 B, in 118 routine patients with gastric cancer, end is followed up a case by regular visits in cut-off, living patients Hp infective dose is starkly lower than Died Patients (P=0.004, t checks), survival group mean ± standard deviation (M ± SD) is 5.9 ± 6.0, and dead group is 8.7 ± 6.4, illustrate that Hp infection level is higher in patients with gastric cancer, survival of patients is poorer.When 50% patient is dead after surgery, namely postoperative 24 months time, survival with Died Patients M ± SD be respectively 7.7 ± 6.4,9.8 ± 6.4, between two groups, when 24 months after surgery, Hp infective dose does not have difference; Within postoperative 24 months, to following up a case by regular visits to end during this period of time, survival group organize M ± SD that Hp infects be respectively 5.9 ± 6.0,9.7 ± 6.5 with dead, and the Hp infective dose that death is organized is apparently higher than survival group (P=0.021).
Survival and Died Patients obvious Hp infect difference and appear at " 0-124 month " and " individual month of 24-124 " patient in these two periods, before illustrating postoperative 24 months, the factor affecting patient's life and death is a lot, comprise the severity of disease, whether have transfer, whether operation is thorough, the general physique situation of patient, the even toxic action of chemicotherapy and the therapeutic response etc. of patient, it is one of them that Hp infects.24 months time, be subject to patient's death all successively of various factors and death.After 24 months, the impact that Hp infects highlights gradually.Hp may as mark, and the effect of the lethal factor relevant to Hp is highlighted out, and Hp may affect existence by interference immune response.Such as, the microenvironment that Hp-Stat3-IL17 is formed, may more be conducive to growth of cancer cells, diffusion, transfer, thus may affect existence.Present design analytical results is pointed out, and Hp infects as mark, may be larger to the Prognostic after 24 months, more should follow up a case by regular visits to interval time by reasonable arrangement.
Patients with gastric cancer follows up a case by regular visits to reference time scope and anti-Hp infects suggestiveness scheme
1.Hp infects negative patient without the need to special follow up time, is undertaken by international guidelines: within every 3-6 month, follow up a case by regular visits to 1 time, 1-2 altogether; Within every 6-12 month afterwards, follow up a case by regular visits to 1 time, 3-5 altogether; Annual 1 time later;
2.Hp infects the radical cure treatment that positive patient recommends to carry out Hp after chemotherapy, the experiment and periodic monitoring C14 exhales, this programme proposes the poor prognosis of Hp infected patient, and relevant with the amount that Hp infects (as Fig. 2 shows), find that patient's death toll accounts for total death toll 82.8% (72/87) in 3 years according to our Follow-up Data, we advise following up a case by regular visits to and should strengthen in postoperative 3 years of patients with gastric cancer, and follow up time should carry out corresponding shortening with Hp infective dose: (1) 1 < Hp < 15, within in postoperative 3 years every 3-4 month, follow up a case by regular visits to once, at least follow up a case by regular visits to 3 times every year, 4-5 follows up a case by regular visits to once every half a year, (2) Hp >=15, follow up a case by regular visits to once, at least follow up a case by regular visits to 4 times every year in postoperative 3 years every 2-3 month, 4-5 follows up a case by regular visits to once (as Fig. 1 shows) for every 4 months.
3. infect positive advising property of patients with gastric cancer Hp eradication therapy for Hp: due to standard triple therapy (PPI+ clarithromycin+amoxycilline Trihydrate bp or PPI+ clarithromycin+metronidazole) eradication rate lower than or far below 80%, the course for the treatment of of this programme also recommended standard triple therapy extended to 10 days or 14 days from 7 days.
It is negative and positive according to Hp that the method for the embodiment of the present invention is suitable for patients with gastric cancer, and the number of infective dose, patient is divided into different grades, formulate different follow up time according to different grades, namely Hp infective dose is higher, and it is shorter to follow up a case by regular visits to interval time, can change by Timeliness coverage conditions of patients, and in patients with gastric cancer, carry out the treatment of anti-Hp infection, likely improve life in patients to a certain extent, extend patient's life-span.Patient for different Hp infection conditions proposes individuation and follows up a case by regular visits to scheme, is made full use of by medical resource, avoids waste, mentions the highest by medical cost performance, therefore, can be individual, country and society saving human cost, material resources cost and financial resources cost in theory.Patient for different Hp infection conditions proposes individuation and follows up a case by regular visits to scheme, can be patients with gastric cancer Timeliness coverage disease progression to the full extent, obtains medical treatment in time, extends survival time, improves the quality of living.
The above; be only the specific embodiment of the present invention, but protection scope of the present invention is not limited thereto, is anyly familiar with those skilled in the art in the technical scope that the present invention discloses; change can be expected easily or replace, all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of described claim.
Claims (10)
1., based on the patients with gastric cancer Prediction of survival method that Hp DNA molecular is quantitative, it is characterized in that, comprise the steps:
1) the relative infective dose of Hp in stomach organization is detected;
2) prognosis evaluation is carried out according to the relative infective dose of the Hp detected;
Wherein prognosis mala degree increases with the rising of the relative infective dose of Hp.
2. method according to claim 1, is characterized in that, the detection of the relative infective dose of Hp adopts real-time fluorescence quantitative PCR.
3. method according to claim 1, is characterized in that, reaction conditions is, 95 DEG C of denaturation 5min, 95 DEG C of sex change 10s, and 60 DEG C of annealing extend 30s, 40 circulations.
4. method according to claim 1, is characterized in that, the relative infective dose of Hp is obtained by following formula, relative infective dose=2 of Hp
-△ △ Ct, wherein △ △ Ct=(Ct sample object-Ct sample internal reference)-(Ct blank group object-Ct blank group internal reference), selects blank group to be negative findings for standard with methylene blue dyeing and regular-PCR; The relative infective dose of blank group is 1, namely≤1 is that HP is negative, and > 1 is that Hp is positive, and experiment repetition is averaged for 3 times.
5., based on the patients with gastric cancer individuation follow up time appraisal procedure that Hp DNA molecular is quantitative, it is characterized in that, comprise the steps:
1) the relative infective dose of Hp in stomach organization is detected;
2) term of reference of the relative infective dose of patients with gastric cancer prognosis mala Hp is formulated;
3) the relative infective dose of the Hp of detection and gained term of reference are compared, determine the scheme of following up a case by regular visits to.
6. method according to claim 5, is characterized in that, described term of reference is as follows: Helicobacter pylori infection negative patient, namely when the relative infective dose of Hp is less than or equal to 1, without the need to special follow up time, undertaken by international guidelines: within every 3-6 month, follow up a case by regular visits to 1 time, 1-2 altogether; Within every 6-12 month afterwards, follow up a case by regular visits to 1 time, 3-5 altogether; Annual 1 time later;
Helicobacter pylori infection positive patient, namely when the relative infective dose of Hp is greater than 1, after chemotherapy, recommend the radical cure treatment carrying out Hp, the experiment and periodic monitoring C14 exhales, and follow up time should carry out corresponding shortening with Hp relative to infective dose:
During 1 < Hp relative infective dose < 15, within postoperative 3 years every 3-4 month, follow up a case by regular visits to once, at least follow up a case by regular visits to 3 times every year; 4-5 follows up a case by regular visits to once every half a year;
Relative infective dose >=15 of Hp, follow up a case by regular visits to once, at least follow up a case by regular visits to 4 times every year in postoperative 3 years every 2-3 month; 4-5 follows up a case by regular visits to once for every 4 months.
7. method according to claim 6, is characterized in that, the radical cure treatment plan of Hp is as follows: adopt standard triple therapy, i.e. PPI+ clarithromycin+amoxycilline Trihydrate bp or PPI+ clarithromycin+metronidazole, the course for the treatment of is 10 days or 14 days.
8. method according to claim 5, is characterized in that, the detection of the relative infective dose of Hp adopts real-time fluorescence quantitative PCR.
9. method according to claim 8, is characterized in that, reaction conditions is, 95 DEG C of denaturation 5min, 95 DEG C of sex change 10s, and 60 DEG C of annealing extend 30s, 40 circulations.
10. method according to claim 8, is characterized in that, the relative infective dose of Hp is obtained by following formula, relative infective dose=2 of Hp
-△ △ Ct, wherein △ △ Ct=(Ct sample object-Ct sample internal reference)-(Ct blank group object-Ct blank group internal reference), selects blank group to be negative findings for standard with methylene blue dyeing and regular-PCR; The relative infective dose of blank group is 1, namely≤1 is that HP is negative, and > 1 is that Hp is positive, and experiment repetition is averaged for 3 times.
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| CN113490752A (en) * | 2018-12-28 | 2021-10-08 | Cj第一制糖株式会社 | Genetic marker for helicobacter pylori related diseases |
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