CN102928424B

CN102928424B - Method and detection kit for detecting cervical fluid heme

Info

Publication number: CN102928424B
Application number: CN201210422484.XA
Authority: CN
Inventors: 温鑫; 崔子伦; 杜晓丽
Original assignee: QINGDAO BELLO BIOTECHNOLOGY CO Ltd
Current assignee: Qingdao Hongliyuan High-tech Co.,Ltd.
Priority date: 2012-10-30
Filing date: 2012-10-30
Publication date: 2014-06-11
Anticipated expiration: 2032-10-30
Also published as: WO2014067033A1; CN102928424A

Abstract

The invention provides a method for detecting cervical fluid heme. The method comprises the following steps of: wiping at the cervical part with little pressure to obtain the fluid; processing the fluid with a tri-hydroxymethyl aminomethane-hydrochloric acid buffer solution with a PH value of 7.4; and then reacting with a color producing reagent, comparing with a shade guide and recording the detection result. The method provided by the invention is simple and practical and low in cost, and can quickly screen the sample without heme, the sample with heme and the sample possibly with heme in the cervical fluid, thereby providing preparation for further screening. The invention also relates to a detection kit for detecting the cervical fluid heme.

Description

A kind of method that detects uterine neck sepage protoheme

Technical field

The present invention relates to a kind of detection method, be specifically related to a kind of method that detects uterine neck sepage protoheme, the invention still further relates to a kind of dedicated test box that detects uterine neck sepage protoheme simultaneously.

Background technology

Protoheme in uterine neck sepage often by the protoheme of histocyte oxygen receptor and FixL transmembrane protein discharge, tumour cell protoheme is separated out or microcirculation is oozed out and caused, the content of its protoheme is few, naked eyes non-availability is shown in, generally need detect by advanced instrument and analytical chemistry method, expense is high, cycle is long, operate complicated, so invention is a kind of simple to operate, detection method with low cost determines in uterine neck sepage whether contain protoheme fast, becomes a problem of needing solution badly.

Summary of the invention

In order to solve, in prior art detection uterine neck sepage protoheme method, expense is high, the cycle long, operate complicated problem, the invention provides a kind of method that detects uterine neck sepage protoheme, of the present invention highly sensitive, with low cost, can detect fast the protoheme in uterine neck sepage; The present invention simultaneously also provides a kind of special detection box that detects protoheme in uterine neck sepage.

Technical scheme of the present invention is as follows:

The method of protoheme in detection uterine neck provided by the invention sepage, comprises following detecting step:

1) bring emgloves, fingerstall on forefinger first knuckle place band, uterus neck a little pressure wiping obtain sepage; Wherein, sepage amount be 0.5 ~ 1ml;

2) fingerstall with sepage is put into sample cup, sample cup is built-in with tris-HCI buffer, and the amount of damping fluid is 3ml, mixes, and obtains sample liquid; Preferably, the pH value of tris-HCI buffer is 7.4;

3) in test pool, order adds sample liquid 1ml, reagent A 0.5ml, reagent B 0.5ml; Mix, in 2 minutes, observe change color, compare and record testing result with color board; The compound method of described reagent A is as follows: get 3,3', it is that 4.4 citrate buffer solution 100ml mix and form that 5,5'-tetramethyl benzidine 50mg is dissolved in PH; The superoxol that described reagent B is 3%.

The present invention provides a kind of detection box that detects uterine neck sepage protoheme simultaneously, the slip lid that comprises box body and be slidably connected with box body, described slip lid is provided with two bar-shaped troughs of outwardly convex, dropper is housed in bar-shaped trough, bar-shaped trough is from sealed bottom, described box body inner hollow, one side is provided with color board, the sample cup rest area that box body inner bottom part is also provided with test pool and places sample cup, test pool side is placed with fingerstall, described test pool extends internally from box body bottom, and middle part is bowl structure to lower recess.

Further, described sample cup rest area is located between color board and test pool.

Further, described color board is parallel with box body bottom, and is positioned at the middle and upper part of box body.

Further, described dropper is with scale.

Further, described sample cup is provided with gland bonnet, and the cup of sample cup is cylindric.

Further, the material of described fingerstall is the nonwoven fabrics of tool adsorbability.

Detection method provided by the invention, simple and practical, with low cost, examination fast goes out in uterine neck sepage the sample containing protoheme, containing the sample of protoheme with may contain the sample of protoheme, for further checking and prepare; The detectable minimum protoheme concentration of the present invention can reach 0.5ng/L, highly sensitive; The present invention simultaneously also provides a kind of detection box, can specially hold vessel and the reagent in testing process, used, this detection box is simple in structure, only needs a detection box just can complete whole detecting step, save space and reagent, also alleviate operator's work burden simultaneously.

Accompanying drawing explanation

Fig. 1 is for detecting box external structure schematic diagram;

Fig. 2 is box body inner structure schematic diagram;

Fig. 3 is fingerstall structural representation;

Fig. 4 is sample cup structural representation;

Wherein: 1. slip lid; 2. bar-shaped trough; 3. sample cup rest area; 4. test pool; 5. color board; 6. fingerstall; 7. sample cup; 8. box body.

Embodiment

Below in conjunction with embodiment, the present invention is described further.

Embodiment 1

a method that detects uterine neck sepage protoheme, method is as follows: slip off the slip lid 1 that detects box, from the interior taking-up fingerstall 6 of box body 8, bring emgloves, then fingerstall 6 put at forefinger the first joint, uterus neck a little pressure wiping obtain sepage, wherein, sepage amount is 0.5-1 ml; The sample cup 7 of sample thief cup rest area 3, inserts pH value and is 7.4 tris-HCI buffer 3 ml, and the fingerstall 6 with sepage is put into sample cup 7, mixes; From two droppers of two interior taking-ups of bar-shaped trough 2, two droppers are preinstalled with respectively reagent A and reagent B, add sample liquid 1ml in the interior order of test pool 4, reagent A 0.5ml, reagent B 0.5ml, mixes, in 2 minutes, observe change color, compare and record testing result with color board 5; The compound method of described reagent A is as follows: get 3,3', 5,5'-tetramethyl benzidine 50mg is dissolved in PH4.4 citrate buffer solution 100ml and mixes and form; The superoxol that described reagent B is 3%.

Testing result is judged as follows:

Negative if do not develop the color, illustrate in sepage without protoheme; If show, light green blueness, to light blue, may contain protoheme, needs further to detect and determine; Blue illustrate and in sepage, contain protoheme to mazarine if show.

For determining the sensitivity of detection method provided by the invention, the sample that shows different colours further to be verified with mass spectrometer, method is as follows:

Get respectively to detect as not developing the color, show that light green blueness is extremely light blue, blue and carry out the detection of protoheme with mass spectrometer to each three examples of navy blue sample, testing result is as follows: that does not develop the color does not all survey and hemochrome; Light green color to light blue is: 0.5 ng/L, 1.7ng/L, 4.3 ng/L; Blueness to mazarine is 20ng/L, 2.5 μ g/L, 52 μ g/L.

Can find out by above data, detection method provided by the invention can make content of hemachrome be less than the sepage colour developing of 1ng/L, sepage to content of hemachrome more than 20ng/L just can be determined and in sepage, contains protoheme, so the sensitivity of detection method provided by the invention is higher, can be used for the protoheme in uterine neck sepage to carry out qualitative detection completely.

Embodiment 2

Below in conjunction with Fig. 1 to Fig. 4, dedicated test box of the present invention is described further.

The detection box of detection uterine neck sepage protoheme provided by the invention, the slip lid 1 that comprises box body 8 and be slidably connected with box body 8, slip lid 1 can slide along box body 8 upper surface directions, described slip lid 1 is provided with two bar-shaped troughs 2 of outwardly convex, a dropper is housed in bar-shaped trough 2, bar-shaped trough 2 is from sealed bottom, box body 8 inner hollow, one side of box body 8 is provided with color board 5, the sample cup rest area 3 that box body 8 inner bottom parts are also provided with test pool 4 and place sample cup 7, test pool 4 limits are placed with fingerstall 6, described test pool 4 extends internally from box body 8 bottoms, middle part is bowl structure to lower recess, described sample cup rest area 3 is located between color board 5 and test pool 4, described color board 5 is parallel with box body 8 bottoms, and be positioned at the middle and upper part of box body 8, described dropper is with scale, described sample cup 7 is provided with gland bonnet, the cup of sample cup 7 is cylindric, the material of fingerstall 6 can be the nonwoven fabrics of tool adsorbability.

Claims

1. a method that detects uterine neck sepage protoheme, is characterized in that, comprises following detecting step:

1) get uterine neck sepage;

2) fingerstall with sepage is put into sample cup, sample cup is built-in with tris-HCI buffer, and the amount of damping fluid is 3ml, mixes, and obtains sample liquid;

3) in test pool, order adds sample liquid 1ml, reagent A 0.5ml, reagent B 0.5ml; Mix, in 2 minutes, observe change color, compare and record testing result with color board; The compound method of described reagent A is as follows: get 3,3', 5,5'-tetramethyl benzidine 50mg is dissolved in PH4.4 citrate buffer solution 100ml and mixes and form; The superoxol that described reagent B is 3%;

The amount of described sepage is 0.5-1ml;

The pH value of described tris-HCI buffer is 7.4.